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. 2018 Nov 5;217(11):3863–3872. doi: 10.1083/jcb.201801208

Figure 5.

Figure 5.

DrrA-mediated Rab1 activity is required for the recruitment of Sec15 to the LCV. (A) HeLa-FcγRII cells were cotransfected with GFP-DrrA61–647 or -DrrA451–647 and 3x-FLAG-Sec15 (wild-type) or -Sec15N691A. 24 h after transfection, cells were fixed and stained with an antibody against FLAG. Arrows indicate DrrA-positive structures. Bar, 5 µm. (B) HeLa-FcγRII cells were transfected with 3x-FLAG-Sec15. 24 h after transfection, cells were infected with wild-type Legionella or ΔdrrA mutant strain for 1 h at MOI 10 and fixed, and extracellular Legionella were stained. After staining, cells were permeabilized and stained with an antibody against FLAG. Intracellular Legionella were detected by Hoechst 33342. Bar, 1 µm. Values are the mean ± SD (n = 3, 100 vacuoles in each experiment). *P < 0.05 compared with ΔdrrA. (C) HeLa-FcγRII cells were cotransfected with GFP-Sec15 and FLAG-Rab1 wild-type or -Rab1N121I. 24 h after transfection, cells were infected with wild-type Legionella for 1 h at MOI 10, fixed, and stained with an antibody against FLAG. Legionella were detected by Hoechst 33342. Bar, 2 µm. Values are the mean ± SD (n = 3, 100 vacuoles positive for Rab1 in each experiment). **P < 0.01 compared with Rab1N121I.