Figure 4.

EGFP-CCDC146 is recruited to the periphery of the inclusion membrane in C. trachomatis infected cells. HeLa cells transfected with plasmids encoding either EGFP-CCDC146_FL or EGFP were left uninfected (UI) or infected for 24 h with C. trachomatis L2/434 (A–D) or C. trachomatis L2/434 harboring pCT288-2HA (E). (A) The cells were fixed with methanol, immunolabeled with anti-GFP and anti-γ-tubulin antibodies, and appropriate fluorophore-conjugated secondary antibodies, and analyzed by confocal immunofluorescence microscopy. The arrows in each panel highlight the γ-tubulin-labeled centrosome. (B) Percentage of uninfected or C. trachomatis-infected HeLa 229 cells showing co-localization between EGFP-CCDC146_FL and γ-tubulin. Data represents three independent experiments (100 cells counted per experiment). P-values were calculated by a two-tailed unpaired Student's t-test relative to UI cells. ^*P < 0.05. (C–E) The cells were fixed with paraformaldehyde 4% (w/v), immunolabeled with anti-MOMP (C), anti-CT442 (D), or anti-HA (E) antibodies, and appropriate fluorophore-conjugated secondary antibodies, and analyzed by confocal immunofluorescence microscopy. All scale bars, 10 μm.