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. 2014 Jul 3;10(7):e1004222. doi: 10.1371/journal.ppat.1004222

Figure 8. The host kinase PI4KIIIα is important for generation of a pool of PI4P used for DrrA targeting during infection.

Figure 8

(A) Influence of knockdown of PI4KIIIα and PI4KIIIβ on DrrA-mediated recruitment of YFP-Rab1b to LCVs in HEK293 cells that stably express FcγR and YFP-Rab1b. After siRNA knockdown for 3 d, the HEK293 FcγR YFP-Rab1b cells were infected with wild type L. pneumophila expressing dsRed for 1 h before fixation. Quantification shown was obtained from at least 3 independent experiments with >300 vacuoles assessed for Rab1 recruitment for each condition. (B) Graph showing the percentage of Rab1b-positive LCVs formed by L. pneumophila strain Lp02+pAM239, expressing GFP, at 30 min post-infection in HEK293 FcγRII cells transiently expressing the human Sac1p phosphatase or the vector control. Data is from three independent experiments with >300 vacuoles assessed for Rab1 recruitment for each condition in each experiment. (C) Quantification of Rab1 colocalization with L. pneumophila wild type or ΔsidF strains, constitutively expressing GFP from plasmid pAM239C, at 60 min post infection in HEK293 FcγRII cells transfected with non-targeting (mock) or siRNAs targeting PI4KIIIα at 3 days prior to infection.