This suggests a stepwise pathway of establishing the mature, fusion-competent chlamydial inclusion. We have shown that inclusion fusion occurs at Crizotinib host cell centrosomes and that in order for fusion to result in a single inclusion, nascent inclusions must be transported by dynein along intact, anchored microtubules to a single site. Comprehending the role of microtubule trafficking in inclusion fusion dynamics is crucial to a complete understanding of the mechanisms by which this obligate intracellular pathogen promotes its intracellular survival and pathogenicity. Electronic supplementary material Additional file 1: Inclusion fusion occurs at minus ends of microtubules. Movie of Figure 1. (M4V 734 KB) Additional file

2: Figure 2: Centrosome positioning affects chlamydial

inclusion localization. Uninfected and infected neuroblastomas were plated on CYTOOchips (glass coverslips imprinted with fibronectin micropatterns). Each micropattern is indicated in the lower left of the top panel. Infected cells were fixed at 12 and 24 hpi (top and bottom panel for each shape, respectively). Cells were stained with antibodies to g-tubulin (green) and Chlamydia (red). Nucleic acid is visualized by staining with DRAQ5 (blue). (TIFF 1 MB) References 1. Weinstock H, Berman S, Cates W: Sexually transmitted diseases among American youth: incidence and prevalence estimates, 2000. Perspect Sex Reprod Health 2004, 36:6–10.PubMedCrossRef 2. Clifton DR, Fields KA, Grieshaber SS, Dooley CA, Fischer ER, Mead DJ, Carabeo RA, Hackstadt T: A chlamydial type III translocated BMN673 protein is tyrosine-phosphorylated at the site of entry and associated with recruitment of actin. Proc Natl Acad Sci USA 2004, 101:10166–10171.PubMedCrossRef 3. Dehoux P, Flores R, Dauga C, Zhong G, Subtil A: Multi-genome identification and characterization of chlamydiae-specific type III

secretion substrates: the Inc proteins. BMC Genomics 2011, 12:109.PubMedCrossRef 4. Hackstadt T, Fischer ER, Scidmore MA, Rockey DD, Heinzen RA: Origins and functions of the chlamydial click here inclusion. Trends Microbiol 1997, 5:288–293.PubMedCrossRef 5. Grieshaber SS, Grieshaber NA, Hackstadt T: Chlamydia trachomatis uses host cell dynein to traffic to the microtubule-organizing center in a p50 dynamitin-independent process. J Cell Sci 2003, 116:3793–3802.PubMedCrossRef 6. Geisler WM, Suchland RJ, Rockey DD, Stamm WE: Epidemiology and clinical manifestations of unique Chlamydia trachomatis isolates that occupy nonfusogenic inclusions. J Infect Dis 2001, 184:879–884.PubMedCrossRef 7. Ridderhof JC, Barnes RC: Fusion of inclusions following superinfection of HeLa cells by two serovars of Chlamydia trachomatis. Infect Immun 1989, 57:3189–3193.PubMed 8. Fields KA, Fischer E, Hackstadt T: Inhibition of fusion of Chlamydia trachomatis inclusions at 32 degrees C correlates with restricted export of IncA. Infect Immun 2002, 70:3816–3823.PubMedCrossRef 9.