Proximity-dependent proteomics of the Chlamydia trachomatis inclusion membrane reveals functional interactions with endoplasmic reticulum exit sites
Fig 4
RNAi validation of inclusion interacting proteins and comparison to previous data sets.
(A) IFU determination following RNAi depletion of 64 proteins identified from early (8 hpi) inclusions. Cells were transfected with siRNA corresponding to targets shown on the y-axis, infected with C. trachomatis L2, and harvested for IFU determination at 48 hpi. Bars mean(SD); n = 3; IFU shown relative to mean IFU of plate. Black squares next to RNAi targets (y-axis) indicate which time points the protein was enriched in APEX2 mass spectrometry data. Teal bars correspond to at least 1.5-fold increase/decrease compared to the mean, blue bars are over 2 fold increase/decrease compared to the mean. Significance was determined by one-way ANOVA with Dunnett’s multiple comparisons test, comparing to mean infectivity of plate; *, p < 0.05; **, p < 0.01; ****, p < 0.0001. (B) Summary of all proteins shared between APEX2 data and two previous inclusion mass spectrometry data sets [15,16]. Lower table describes all proteins that overlap between APEX2 and AP-MS Inc-specific interactions [15]. Shaded boxes represent significant enrichment found by APEX2. (C) Summary of APEX2 identified proteins with reported microscopy-based associations with Chlamydia inclusions. PMID entries refer to the publications used to provide this evidence.