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

Figure 3.

Figure 3.

Expression of mutant Sec15 inhibits recruitment of Sec22b to the LCV. (A) HEK293-FcγRII cells were cotransfected with plasmids encoding GFP, GFP-DrrA61–647 or -DrrA451–647, and FLAG or FLAG-Rab1A. 24 h after transfection, cell lysates were prepared and immunoprecipitated (IP). (B) HEK293-FcγRII cells were transfected with plasmids encoding 3x-FLAG-Sec15 (wild-type) or -Sec15N691A and GFP-Rab1. 24 h after transfection, cells were infected with wild-type Legionella for 1 h at MOI 50. After infection, cell lysates were prepared and immunoprecipitated. (C) HEK293-FcγRII cells were cotransfected with 3x-FLAG-Sec15 (wild-type) or -Sec15N691A and RFP-Rab1 or -Sec22b. 24 h after transfection, cells were infected with wild-type Legionella for 1 h at MOI 5, fixed, and stained with antibodies against FLAG and Legionella. Localization of RFP-Rab1 and RFP-Sec22b was assessed for the vacuoles containing wild-type Legionella. Values are the mean ± SD (n = 3, 50 vacuoles in each experiment). n.s., not significant. *P < 0.05 compared with wild-type. Bar, 10 µm.