FIG 5.

Chlamydia ABs differ in their presentation of the mid-stage inclusion membrane protein IncA. HeLa cells were infected at an MOI of 1. Cells were either left untreated or treated with various aberrance inducers, and cells under all conditions were fixed at 24 hpi. Immunolabeling was conducted for IncA as well as MOMP. (a) Ampicillin (AMP) and DPP treatments. See Fig. S4 in the supplemental material for the rest of the conditions. Images are representative of between 10 and 20 inclusions observed under each condition, and the experiment was carried out three times. (b) To assess the fusogenic potential of inclusions under aberrance induction, HeLa cells were infected at an MOI of 10, fixed at 24 hpi, and labeled for CT229. Inclusions present per cell were measured by counting cell and inclusion numbers across 10 random imaging planes under each condition (untreated, AMP treatment, and DPP treatment). Data points represent average values of the numbers of inclusions per cell per imaging plane examined with 10 imaging planes under each condition. The assay was carried out at least twice under each condition tested. Lines represent mean values for all data points under each condition, and error bars represent standard deviations. **, P < 0.005; ns, not significant. (c) Representative images from the analysis of fusogenic and nonfusogenic inclusions. The rightmost panels are magnifications of the boxed areas. Bars, ∼2 μm.