Acquisition of Rab11 and Rab11-Fip2—A novel strategy for Chlamydia pneumoniae early survival
Fig 2
Development of early C. pneumoniae inclusion is dependent on Akt/PIKfyve activity.
(A) Quantification of colocalization of Rab7 and EGFR with chlamydial EBs during the first hour p.i. in cells transiently transfected with GFP-Rab7 (n = 3). (B) Confocal images of GFP-Rab7 colocalizing with EBs in PI3P-positive endosomes (visualized by mCherry-2xFYVE and DAPI, respectively) at 15 min (top row) and 30 min p.i. (bottom row). White arrows indicate colocalization. Bar 1 μm. (C–E) Quantification of EB internalization in cells pretreated with the indicated inhibitor for 2 h prior to infection. Internalization was analyzed in 30 individual cells. (C) Entry of EBs into cells pretreated with the Akt inhibitor MK22 (3 μmol) or DMSO for 2 h prior to infection. Internalization was quantified microscopically at 2 h p.i. (n = 3). (D) Effects of pretreatment with the PIKfyve inhibitor YM201636 (800 nmol) for 2 h prior to infection on EB internalization (n = 3). (E) Quantification of GFP-Rab7 colocalization with EBs in PI3P-positive endosomes (visualized by mCherry-2xFYVE and DAPI) at 30 min p.i. in cells pretreated with MK22, the PIKfyve inhibitor or DMSO (n = 3). *** P value ≤0.001, n.s. P value <0.05.