VEGF (NM_001025250.2) forward: 5′-CCA CGA CAG AAG GAG AGC A-3′ and reverse: 5′-AAT CGG ACG GCA GTA GCT T-3′ 80 bp. IL-6 (NM_031168.1) forward: 5′-TCT CTG GGA AAT CGT GGA www.selleckchem.com/products/Docetaxel(Taxotere).html A-3′ and reverse: 5′-TCT GCA AGT GCA TCA TCG T-3′ 81 bp. IL-1β (NM_008361.3) forward: 5′-GTT GAC GGA CCC CAA AAG-3′ and reverse: 5′-GTG CTG CTG CGA GAT TTG-3′ 93 bp. IL-10 (NM_010548.1) forward: 5′-TCCCTGGGTGAGAAGCTG-3′ and reverse: 5′-GCTCCACTGCCTTGCTCT-3′ 91 bp. Caspase-3 (NM_009810.2) forward: 5′-TAC CGG TGG AGG CTG ACT-3′ and reverse:
5′-GCT GCA AAG GGA CTG GAT-3′ 104 bp. TGF-β (NM_021578.2) forward: 5′-ATA CGC CTG AGT GGC TGT C-3′ and reverse: 5′-GCC CTG TAT TCC GTC TCC T-3′ 77 bp. HGF (NM_010427.3) forward: 5′-GCC AGA AAG ATA TCC CGA CA-3′ and reverse: 5′-CTT CTC CTT GGC CTT GAA TG-3′ 197 bp. 36B4–Rplp0 (NM_007475.5) forward: 5′-CAA CCC AGC TCT GGA GAA AC-3′ and reverse: 5′-GTT CTG AGC TGG CAC AGT GA-3′ 150 bp. The normality of the data (Kolmogorov–Smirnov test with Lilliefors’ correction) and the homogeneity of variances
(Levene median test) were tested. If both conditions were satisfied, differences between the Sham and CLP groups at day 1 were assessed by two-way ANOVA followed by Tukey’s test. Since no difference was observed between Sham-SAL and Sham-BMDMC at days 1 and 7 we decided to present only one time point. The comparison between CLP-SAL and CLP-BMDMC groups at days 1 and 7 was performed using one-way ANOVA or one-way ANOVA on ranks for parametric and non-parametric data, respectively. Survival curves were derived by the Kaplan–Meier method and compared by log rank test. Data are presented as mean ± standard error of selleck compound mean or median (25th–75th percentiles) as appropriate. A p value < 0.05 was considered statistically significant. Statistical analyses were done with SigmaStat 3.1 (Jandel Scientific, San Rafael, CA, USA). The following subpopulations were identified from the pool of intravenously injected BMDMCs characterized by flow cytometry: total
lymphocyte (CD45+/CD11b−/CD29−/CD34− = 4.2%), Staurosporine cost T lymphocyte (CD45+/CD3+/CD34−=2.1%), T helper lymphocyte (CD3+/CD4+/CD8− = 0.5%), T cytotoxic lymphocyte (CD3+/CD4−/CD8+ = 1.6%), monocytes (CD45+/CD29+/CD14+/CD11b−/CD34−/CD3− = 2.8%), neutrophils (CD45+/CD11b+/CD34−/CD29−/CD14−/CD3− = 78.7%), hematopoietic progenitors (CD34+/CD45+ = 0.5%), and other progenitors cells (CD45− = 9.1%). At day 7, the survival rate of Sham-SAL and Sham-BMDMC mice was 100%. All animals from the CLP-SAL group died within 48 h after sepsis induction. Therefore, we were unable to provide data for CLP-SAL group at day 7. Survival at days 1 and 7 was higher in the CLP-BMDMC compared to CLP-SAL group (75% vs. 60% and 70% vs. 0%, respectively, P < 0.001) ( Fig. 2). Est,L was higher in CLP-SAL animals compared with Sham-SAL at day 1. BMDMCs led to a significant reduction in Est,L at day 1, whereas at day 7 this reduction was more pronounced ( Fig.