FIG 8.

Analysis of C. trachomatis infection in AHNAK-deficient MEFs. (A) WT or ahnak homozygous knockout (KO) MEFs were infected at an MOI of 1 with WT C. trachomatis. Cultures were fixed at 24 h postinfection for direct IFU counts. (B) WT KO MEFs were infected for 1 h at 4°C with C. trachomatis L2 at an MOI of 10. Infections were carried out in the presence of vehicle control or cytochalasin D (CytoD). Cultures were shifted to 37°C for 30 min and then paraformaldehyde fixed. External EBs were specifically labeled with MOMP-specific antibodies, and internalized bacteria were labeled in subsequently permeabilized cultures using Chlamydia-specific antibodies (**, P < 0.0001). (C) KO MEFs were infected with equal particles of WT or tmeA mutant C. trachomatis at an MOI of 0.5. Cultures were processed for direct enumeration of inclusions at 24 hpi (*, P < 0.003). All data are represented as means with standard deviations calculated from triplicate cultures. Student's t test with Welch's correction was employed to assess statistical significance.