Figure 4.

The C. burnetii T4SS effector AnkF is dispensable for CCV maturation. (A, B) HeLa cells were infected with wild-type C. burnetii (WT) and the ankF::Tn (ankF::Tn) mutant at an MOI of 50. At 24 h post-infection the cells were fixed and stained with an anti-LAMP2 antibody (red) and with antisera against C. burnetii (green). Nuclei and bacterial DNA were stained with DAPI (blue). Cells were visualized using LSM. (A) Representative LSM images are shown. (B) 100 infected cells were analyzed for the association of LAMP2 with the CCV. Error bars represent mean standard deviations of three independent experiments. (C, D) HeLa cells were infected with wild-type C. burnetii (WT) and ankF::Tn (ankF::Tn) at an MOI of 50. Cells were incubated with DQ-Red BSA for 16 h. At 24 h post-infection, the cells were fixed and stained with antisera against C. burnetii (green). Nuclei and bacterial DNA was stained with DAPI (blue). Cleaved DQ-Red BSA emits fluorescence (red). Cells were visualized using LSM. (C) Representative LSM images are shown. (D) 100 CCVs were analyzed for red fluorescence using an epifluorescence microscope. Error bars represent mean standard deviation of three independent experiments.