This requirement was subsequently linked to electron transport along with the possession with the sodium pumping respiratory NADH dehydrogenase Nqr. On top of that to electron transport, it’s also been reported that sodium is needed for amino acid transporters and for your oxidation of compounds this kind of as alanine and galactose in some marine bacteria. The ionic specifications of marine bacteria also can consist of calcium and magnesium, but the genetic basis for these requirements is unknown. At current, it remains unclear if very similar marine adaptations take place in Gram beneficial taxa. The discovery of your sodium pumping NADH dehy drogenase Nqr plus the associated genes nqrA F represented the very first genetic hyperlink to sodium dependence in Gram adverse marine bacteria.
Nqr is considered one of 3 varieties of respiratory NADH dehydrogenases and is acknowledged to come about in lots of Gram adverse marine bacteria and some clinical pathogens. When present, Nqr does not preclude the occurrence of other NADH dehydro genases within a genome. The far more widespread prokaryotic selelck kinase inhibitor NADH dehydrogenase would be the proton pumping NDH one, that is also known as complex I. NDH one is com posed of 14 genes and displays no homology with Nqr but both are power coupling enzyme com plexes that generate an ionic motive force utilised to produce ATP and drive other cellular processes. Interestingly, the membrane bound, ion pumping nuo genes show considerable sequence similarity to the 6 genes that make up the multi subunit NaH antiporter Mrp. The third form of NADH deydrogenase is NDH 2, and that is generally composed of 1 to some proteins and it is not an power coupling complicated or been linked to marine adaptation.
The capacity of bacteria to adapt to external changes while in the osmotic environment is fundamental to survival. Osmoadaptation in bacteria generally entails the intra cellular accumulation of compatible solutes this kind of as gly cine and betaine. These compounds are acquired either by selleck de novo biosynthesis or straight through the environ ment. Bacteria also have mechanisms to survive osmotic down shock that normally involve a mixture of speci fic and non certain mechanisms of solute efflux along with aquaporin mediated water efflux. A single essential mechanism of solute efflux is mediated from the mechano sensitive channel of big conductance. This membrane bound, stretch activated channel is common in bacteria and believed to act as an emergency value to release turgor stress following sudden osmotic down shock.
From the marine halophile Vibrio alginolyticus, the introduction of mscL alleviated cell lysis following osmotic downshock and thus the product of this gene might signify a crucial mechanism to survive the transition from marine to freshwater environments. On top of that to particular ionic demands and mechanisms to survive osmotic stress, comparative genomics continues to be utilized to identify other mechanisms of marine adaptation.
The down regulation of genes in central metabolic pathways may perhaps reflect the growth limi tation observed during the HacACA mutant. Popular and distinct features from the constitutive activation of HacA and the UPR induction by chemical substances or heterologous protein expression To gain a broader overview with the influence of the cons titutive activation of HacA on the. niger we compared our information set with the data of Guillemette and co staff during which the genome broad transcrip tional protein secretion related stress responses was analyzed. Within this examine, transcriptional targets from the UPR pathway had been recognized by treatment method of the. niger using the ER disturbing chemical agents tunicamycin and dithiothreitol and employing a strain generating the recombinant tissue plasminogen activator as being a model for heterologous protein manufacturing.
As shown in Figure 5, within the induced set of genes, 13 genes are com monly unregulated in the two studies and 81 genes are differentially expressed in HacACA 1 HacAWT in at the very least two in the 3 situations per formed by Guillemette et al. These 94 usually induced genes include all the genes recognized inside the Guillemette et al. research linked to protein folding, i thought about this trans locationsignal peptidase complex and glycosylation and nearly all of the genes that belong on the classes of vesicle trafficking and lipid metabolism. Even so, more genes belonging to each of those cat egories are recognized while in the HacACA 1HacAWT comparison. One of a kind genes located in not less than two from the ailments tested and never in our data set relate largely for the classes of cellular transport, pressure relevant, amino acid metabolic process, carbohydrate metabolic process and unclassified genes.
Vicriviroc For the repressed set of genes we identified 45 widespread genes to our review and Guillemette et al. that are evenly distributed throughout the categories established from the authors. The truth that the quantity of frequently down regulated genes is smaller between the two studies suggests critical variations and heterogeneous responses for the induction on the UPR indirectly along with the manipulation in the transcription element that regu lates this pathway while in the total cell metabolism. The constitutive activation of HacA triggers the induction of ERAD genes Secretory proteins that fail to fold effectively commonly accu mulate while in the ER and therefore are sooner or later targeted to destruction through the proteasome, a method termed ER associated degradation.
Genes encoding proteins that are putatively concerned in ERAD have already been recognized while in the A. niger genome as well as the expres sion of those genes was examined during the microarray information set. As highlighted in Table two, the expression of numerous putative ERAD elements was induced during the HacACA mutant. For instance, the der1 homologue, involved in transport of unfolded proteins from the ER, is 4.
Regulation of Sox9 and NFB activity by TNF are independent of MEKERK signalling We next wanted to determine the possible molecular basis for TNF modulated, U0126 sensitive gene expression. First, we investigated whether U0126 affected the ability of TNF to regulate the activity of the transcription factors Sox9 and NFB, which are known to be regulated by TNF in chondro cytes. As expected, TNF significantly reduced the level of Sox9 activity and increased the level of NFB activity in chondrocytes. There was no signif icant effect, however, on the level of inhibition or the induction of Sox9 and NFB activity, respectively, by either U0124 or U0126. Furthermore, we found that TNF induced DNA binding of NFB was reduced by pre treatment with DMSO and was not further reduced by pretreatment with U0124, U0126 or the selective epidermal growth factor receptor inhibitor, PD153035.
These results indicate that transcrip tion factors other than Sox9 and NFB are targets of TNF induced MEKERK signalling. Egr 1 DNA binding is increased in a TNF induced MEK ERK dependent manner read this post here To determine additional, candidate transcription factors that may regulated by MEKERK, we considered that Egr 1 is a known early target of MEKERK signalling and that IL 1 induc tion of Egr 1 inhibits the activity of the human type II collagen proximal promoter. We therefore focused the remainder of our study on Egr 1 and its possible role in regulating U0126 sensitive TNF induced genes. We identified multiple putative Egr 1 binding sites in the pro moter regions of the rat Col2a1 and Agc1 genes that were proximal to the transcription initiation site and overlapped with putative Sp1 binding sites.
TNF treatment of chondrocytes selelck kinase inhibitor over 24 hours did not alter the Egr 1 protein lev els, and neither did treatment for 90 minutes alter the nuclear localization of Egr 1. We then used electrophoretic mobility shift assays to investi gate whether the binding of Egr 1 to DNA was dependent on TNF induced MEKERK signalling. Nuclear extracts from chondrocytes treated with TNF for 90 minutes increased the DNA binding of two complexes containing Egr 1 to an Egr consensus DNA binding site. Both complexes were reduced when extracts were preincubated with a 100 fold molar excess of double stranded cold Egr con sensus ODNs, but not with cold mutant Egr ODNs or NFB consensus ODNs. Compared with pre incubation of extracts with the anti NFB p65 antibody, prein cubation of extracts with the anti Egr 1 antibody specifically reduced the DNA protein complexes attributed by the Egr consensus ODN competition studies to be a result of Egr DNA binding. Pretreatment of cels with U0126 attenuated the increase in complex formation of both identified complexes. l
Each group consisted of eight or 9 animals. Examination of gene expressions in mouse full hind limbs Instantly just after the animals have been killed, total hind limbs had been immersed in RNAlater RNA Stabilization Reagent and stored at 80 C until eventually total RNA extraction. The entire hind limbs have been excised, quickly soaked in TRIzol, and crushed in the bead mill. Complete RNA was extracted working with an RNeasy kit according to your kit suppliers protocol. Quantities of total RNA obtained from just one complete hind limb have been 4. 5 to 18 ug, 15to 60 ug, 9 to 60 ug, 12 to thirty ug, and four. two and 30 ug. cDNA was synthesized with an Omniscript RT kit making use of random 9 mer primers in accordance on the kit makers protocol.
Quantitative genuine time polymerase chain response was carried out by running a TaqMan gene expression assay, tar geting mouse SLC19A1, IL six, TNF a, and glyceralde hyde three phosphate dehydrogenase, on an ABI PRISM 7500 program in accordance to the manufacturers protocol. Analysis of gene expressions in mouse immune cells For CD4 T cell and B cell subset selleck sorting, splenocytes were labeled with antibodies to CD4 and B220 and sorted to 97% purity through the use of a fluorescence activated cell sorter. Complete RNA was extracted implementing an RNeasy kit according on the kit makers protocol. Synthesis of cDNA and measurement of mRNA levels by quantitative real time PCR were carried out from the very same procedures as described above. Isolation and culture of mouse synovial cells Arthritic mice have been killed and the synovial tissues removed from their hind limbs. Synovial tissues have been incu bated at 37 C for 180 min in a MEM supplemented with 10% fetal bovine serum and containing 0.
5 mgmL of Liberase Blendzyme2. Soon after incubation within the synovial tissues with the Liberase Blendzyme2, the resulting cells have been cultured within a culture flask in a MEM supplemented with 10% FBS along with the non adherent selleck chemicals cells had been removed and discarded. Syno vial cells have been then subcultured in a MEM supplemented with 10% FBS to a density of 2105 cells35 mL in the T175 flask. On this research, synovial cells from passages two to 5 were used. Evaluation of gene expressions in synovial cells Synovial cells were cultured within a MEM supplemented with 10% FBS for 24 h. Following this pre culture, cells had been cultured with mouse IL 6 and soluble mouse IL 6R, TNF a, or MTX for 24 h. Total RNA was extracted working with an RNeasy kit in accordance towards the kit makers protocol. cDNA was synthesized with an Omniscript RT kit making use of random 9 mer primers in accordance to the kit suppliers protocol. Quantita tive true time PCR was carried out by running a TaqMan gene expression assay, focusing on mouse SLC19A1, multi drug resistance protein one, breast cancer resis tance protein, and GAPDH, on an ABI PRISM 7500 procedure in accordance towards the companies protocol.
Irrespective of whether augmented ectodomain shedding of membrane receptors displays greater turnover andor activation from the re spective pathways or rather contributes to their dysfunc tion andor inhibition remains for being clarified. Tactics Participants Plasma samples from 107 SLE individuals participating within the Oklahoma Cohort for Rheumatic Illness had been stud ied. All sufferers satisfied at the very least four on the 1982 revised American Rheumatism Association criteria for SLE. Forty 5 of those individuals had been matched to healthy controls by age, gender and ethnicity. Clinical and labora tory data had been registered right into a database which integrated no personal identifiers. The characteristics from the pa tients and also the controls enrolled are reported in Table 1. Heparinized plasma samples have been collected and stored at 70 C without delay after assortment.
Sickness exercise was scored utilizing the SLEDAI and also the British Isles Lupus Assessment Group index. Levels of com plement fractions C3 and C4 had been established by immu noturbidity. Complete 50% hemolytic complement action was calculated by using a liposome immunoassay. Antinuclear antibodies were detected by indirect im munofluorescence. order MK-1775 The Crithidia luciliae check was used for detection of anti double stranded DNA. Antiextractable nuclear antigen autoantibodies were measured by Ouchterlony double immunodiffusion. Before participation, all participants gave their informed consent to donate their blood samples. The research was approved by the institutional evaluate boards of your Okla homa Healthcare Analysis Basis and Temple University.
Cell cultures Monocytes from buffy coats of wholesome blood donors had been isolated with Ficoll Paque PLUS gradient and by magnetic separation utilizing a kit for human monocyte enrichment by adverse selection ac cording for the manufacturers instructions. The purity of CD14 cells was 90% NVPBHG712 as assessed by flow cytometry. CD14 cells had been cultured for 3 days at 0. 8106 cellsml in 24 properly plates containing serum free of charge X VIVO 15 medium while in the pres ence or absence of macrophage colony stimulating issue. granulocyte macrophage colony stimulating element ,IL ten, IFN. IFN B, IFN, IL 4, IL 17 or dexamethasone. When specified, on day 2, cells had been coin cubated with lipopolysaccharide to the remaining 24 hrs. Cells were then harvested by centrifugation. Supernatants were collected and promptly stored at twenty C to get a couple of days in advance of remaining examined by enzyme linked immunosorbent assay. Pellets had been resuspended in phosphate buffered saline and right away analyzed by flow cytometry. Enzyme linked immunosorbent assay Plasma concentrations of sAxl, sMer and sCD163 were measured by sandwich ELISA according to standard procedures.
Although ERKs are believed to play a essential function in cell prolif eration, it’s been suggested that persistent activation of ERKs might possibly lead to cell cycle arrest and differentiation in PC12 cells, or cell growth inhibition and apoptosis in nor mal rat hepatocytes and numerous human tumor cell lines. On the other hand, a comprehensive time course and transient ERK activations, interspersed with phosphatase mediated inac tivations, are seldom addressed. We and some others showed that the serinethreonine phos phatases PP1 and PP2A participate in the dephosphoryla tion of ERKs. It’s exciting that OA was far more useful in mERhigh cells, whereas cyclosporin A was essentially equally effi cient in each cell populations. These data recommend that the level of mER expressionactivation is usually associated with activation of different kinds of ERK modulating phos phatases.
Other phosphatases, this kind of as tyrosine phos phatase and dual specificity phosphatases , are already implicated in ERK dephosphorylation and will be the subject of our potential scientific studies. More scientific studies of ERK activations, deactivations, and stability will probably be essential in advance of we will formulate a even more glo bal picture from the inhibitor OTX015 submit translational modifications that cause perform of this necessary group of regulators in prolifer ation and differentiation. Yet, it’s clear that nong enomic estrogen actions as well as membrane receptors by way of which they act take part in this regulation. Conclusion E2 induced alterations in breast cancer cell amount is usually straight associated to ERK12 activationdeactivation pat terns and interacting signaling mechanisms.
The differential conduct of cell lines expressing different ranges of mER suggests a function for this receptor while in the temporal coordination of phosphorylationdephosphorylation occasions affecting the mitogen activated kinases ERK1 and ERK2. Introduction Breast cancer is definitely the most normally diagnosed form of cancer as well as the second selelck kinase inhibitor top rated lead to of cancer death in Western girls. Between a single from eight and one particular from ten girls will create breast cancer in the course of her lifetime, together with the sickness becoming a main cause of mortality in girls over the age of 35 many years. Survival rates of individuals with early breast cancer from the Uk and during the Usa have improved steadily in excess of the past 15 years, largely as the outcome of advances in and enhanced entry to early diagnosis and more effective therapy.
More gains, on the other hand, will require new pre ventative and therapeutic strategies that call for considerably better understanding on the genetics and biology of human breast cancer. Such practical knowledge, that’s swiftly accruing since the consequence of postgenomic technologies this kind of as professional teomics and transcriptional profiling, needs to be translated into a setting during which likely clinical responsiveness will be evaluated.
Inter estingly, VCP, a companion of derlin 1 inside the retrotranslocation complicated, was overexpressed in colorectal carcinomas. The VCP expression level is definitely an independent prognosticator for recurrence of colorectal carcinoma and patient survival. Also, preceding research have demonstrated that other strain responsive proteins, which are regulated by unique branches from the UPR, are often overexpressed in particular forms of tumors. In light in the molecular heterogeneity of cancer cells, these findings indicate that a broad transcriptional system induced in the course of the UPR could be extra relevant to cancer than the uncomplicated expression of one of these tension responsive pro teins. Hence, strain proteins may well prove to be useful tumor markers and achievable therapeutic targets.
Conclusion In summary, this study demonstrated that derlin 1 expression will not be constitutively overexpressed in some breast cancer cell lines but is usually substantially induced by serum starvation and agents that disturb ER function. These information suggest that der lin selleckchem 1 expression may be induced by the ER stress that is certainly present in breast cancer. In addition, this study demon strated that derlin 1 knockdown in breast cancer cells rendered cancer cells far more susceptible to ER anxiety induced apoptosis, indicating that derlin 1 overexpression in breast cancer may possibly improve cancer cell survival following exposure to anxiety. General, these findings reveal the involvement of derlin 1, a important part of the UPR, in tumorigenesis by way of improved expression as well as the capability of relieving pressure induced apoptosis.
Introduction Homeobox genes are an essential class of master regulatory genes that encode transcription things responsible for orchestrating developmental processes in many species of animals, as well as in plants and fungi. These genes are char acterized by a conserved 180 nucleotide kinase inhibitor peptide synthesis price sequence coding for any 60 amino acid homeodomain that directs binding to down stream target genes that could be activated or repressed. An rising variety of investigations assistance the involvement of homeobox genes in tumorigenesis of prostate, lung, renal, ovarian, colorectal, and breast tissues. Especially in breast cancer, altered levels of many homeobox genes are straight connected with cellular transformation, disruption in the cell cycle, apoptosis, and progression to a metastatic phe notype. Beta Protein 1 belongs towards the Distal significantly less subfamily of the homeobox gene family members. BP1 maps to chromosome 17q21 22, a area of DNA that’s generally amplified in breast cancer and that contains the tumor suppressor gene BRCA1 along with the oncogene ErbB2. We’ve got identified that BP1 is expressed in 81% of invasive ductal breast tumors.
Following 6 h at 37 C within a humidified 5% CO2 incubator, the cells had been placed in fresh serum containing medium. Cells had been examined just after 48 h within the humidified incubator. Flow cytometry and cell viability To measure cell death, cells had been resuspended in annexin V binding buffer, incubated with five uL of propi dium iodide and subjected to flow cytometry evaluation, making use of a FACS Canto II Flow Cytometer. To allow selective ana lysis in the cells that had incorporated the various hpdODNs, fluorescein labelled hpdODNs had been used. Fluorescein labelled cells were analyzed for PI incor poration or annexin V labelling. A cell death index was established by means of computation of averages. Gel, western blotting Cells were washed in Phosphate Buffered Saline, lysed in sodium dodecyl sulfate sample buffer, 2% SDS, 20% glycerol, 1 mM sodium vanadate, 1 mM dithiothrei tritol and 0.
01% bromophenol blue sonicated and stored at 70 C. Proteins had been separated on SDS polyacrylamide gels and transferred onto nitrocellulose membranes, membranes blocked with dry skimmed milk in Tris Buffered the original source Saline were incubated with antibody overnight at 4 C. Anti phospho STAT1, anti STAT1 and anti STAT3, anti cyclin D1 and anti IRF1 have been employed. Blots had been washed in TBS with Tween, incubated with peroxidase coupled goat anti mouse or goat anti rabbit secondary antibody, washed in TBS T and revealed by chemiluminescence and autora diography. When required, membranes have been stripped with Blot Restore Kit and reprobed with anti tubulin or anti actin antibody to make sure equal loading from the gels. Prestained molecular weight stan dards had been used.
Oligodeoxynucleotide pull down For in cell hpdODN pull down assays, cells have been trans fected together with the biotinylated hpdODNs, as described under transfection, and after that masitinib molecular weight lysed in cell lysis buffer containing salmon sperm DNA. Protein concentration was measured inside the samples. Extracts were recovered on avidin sepharose beads, beads have been incubated for 30 min at 4 C in binding buffer. Soon after washing with binding buffer, complexes had been eluted in SDS sample buffer, separated on SDS Web page, and subjected to immunoblotting using anti STAT1 or anti STAT3 antibodies and processed as above. Immunocytochemistry Cells had been grown at 50 60% confluence in 8 well plates to a density of 105 cells ml. Cells were transfected with fluorescein labelled hpdODNs, incubated, washed in PBS, fixed with 3. 7% formaldehyde for 15 min, permeabilized in 0. 1% Triton X one hundred for 15 min and incubated in 5% FCS 0. 1% Tween PBS for 1 h. Cells have been stained with anti STAT3 or anti STAT1 antibody for two h, then stained with an Alexa fluor 546 labeled secondary antibody for 90 min. Cells, counter stained with four, six diamidino 2 phenylindole, had been mounted onto glass slides with Vectashield.
Methanol and PBS were used as a car control for digoxin and ouabain respectively, during the proliferation assay. Cell proliferation was assessed making use of an alamarBlue assay. U 87, D54 and NTAs had been plated in black clear bottom 96 properly plates and incubated overnight. The following day, each drug was added at its made concentration with 20 ul of 10X alamarBlue reagents. The volume in each and every well was created as much as 200 ul with the development medium. After 72 hours incubation, alamarBlue fluorescence was mea sured on a Perkin Elmer Wallac 1420 Multilabel counter using a 540 nm excitation filter in addition to a 590 nm emission filter. Fold inhibition was calculated by dividing the fluorescence values for manage cells with fluorescence values of cells treated using a particular concentration of cardiac glycosides.
For apoptosis analysis, both U 87 and NTAs had been plated within a six well plate and incubated for 24 hours. Immediately after 24 hours, cells had been trea ted with 500 nM of digoxin and ouabain overnight Omecamtiv mecarbil clinical trial and observed below a microscope Statistical Evaluation GraphPad Prism 5 software program was applied to compute all of the survival curves. To establish the clinical outcome, patient survival was used as a measure where survival was defined because the time in days from 1st surgical resec tion of GBM to death. Out of 21 patient samples, survi val data had been readily available for 16 distinctive sufferers. Outcomes Mutations in Sodium Ion Channels are Related with Shorter Survival in GBM Individuals Systematic analyses of functional gene groups and path techniques from a earlier study identified ion channel genes that transport sodium, potassium or calcium ions as among the gene groups most frequently mutated in GBM.
The sodium, potassium and calcium ion transport gene groups were every single evaluated to determine if muta tions in these gene groups altered average patient survi val. Nineteen in the 21 individuals showed selleck at the very least 1 mutation in sodium, potassium or calcium channels taken collectively. Fourteen sodium channel genes, 13 potassium channel genes and 18 calcium channel genes had somatic mutations. None of the mutations have been identified in a lot more than 1 patient except for SCN9A, CACNA1H, and TRPV5 exactly where every gene was mutated in two individuals. Interestingly, each of the samples with IDH1 mutations didn’t have any sodium channel mutations. A comprehensive list of genes was divided into person lists of genes that were associated with sodium channels, potassium channels or calcium channels.
As an example, sufferers have been classified in to the sodium channel mutation group if they had a mutation in no less than 1 sodium channel gene. If there have been no mutations in any sodium chan nels, the patients have been grouped into a sodium channel unmutated group. Patients had been grouped inside a equivalent way for potassium channels and calcium channels.
Adhesion assay HRMCs had been grown to confluence in six nicely plates with coverslips, incubated with LPS for 16 h, after which adhe sion assays had been performed. Briefly, THP 1 cells have been labeled having a fluorescent dye, 10 uM BCECF AM, at 37 C for 1 h in RPMI 1640 medium and subsequently washed with PBS followed by centrifuga tion. Confluent HRMCs in six effectively plates had been incubated with THP 1 cells at 37 C for 1 h. Non adherent THP 1 cells had been removed and plates have been gen tly washed twice with PBS. The numbers of adherent THP 1 cells had been determined by counting four fields per 200X high energy field effectively applying a fluorescence microscope. Experiments have been performed in triplicate and repeated at the very least 3 instances.
Co immunoprecipitation assay Cell lysates containing 1 mg of protein have been incubated with two ug of an anti c Src or anti p300 antibody at 4 C for 24 h, and after that ten ul of 50% protein A agarose beads was added and mixed at 4 C for 24 h. The immunoprecipitates selleck inhibitor had been collected and washed thrice using a lysis buffer with out Triton X 100. 5X Laemmli buffer was added and sub jected to electrophoresis on SDS Web page, and after that blotted working with an anti TLR4, anti p47phox, anti c Src, anti p300, or anti ATF2 antibody. Evaluation of information Information had been estimated applying a GraphPad Prism System. Quantitative data have been expressed as the indicates SEM and analyzed by one way ANOVA followed with Tukeys post hoc test. P 0. 05 was deemed considerable. Lay abstract Aggressive Non Hodgkin lymphomas are a het erogeneous group of lymphomas derived from germinal centre B cells. 30% of NHL sufferers do not respond to treatment.
Present criteria to distinguish person NHL subtypes including morphology, immunophenotype, and genetic abnormalities do not permit trusted subtype categorization and prediction of treatment response for NHL instances. The pathological mechanisms behind this heterogeneity are poorly understood. Hence there is a need of new and more solutions discover more here for stratifying NHL. The goal of our research will be to estimate the extent to which distinct signal transduction pathways could possibly be re sponsible for the variations in gene expression that distin guish individual lymphomas. We postulate that signals linked with all the immune response can resemble path strategies activated in distinct NHL subtypes.
To get closer insight in to the relevance of distinct cell signaling networks to NHL subtypes, we stimulated human transformed germinal centre B cells with components known to modify B cell signalling, or that are involved in B cell microenvironment or lymphoma pathogenesis. We discov ered that coherent gene expression patterns, associated to dis tinct in vitro stimuli, characterize individual NHLs. Exemplified by an IgM stimulation we identified signal ling pathways dominantly involved in regulating this con sistent global gene expression pattern.