Ectopic expression of WT MTMR 4 restores the effects of MTMR 4 loss. (A and B) T47D cells co-expressing MTMR 4 shRNA66 and GFP-KRASG12V (A) or GFP-LactC2 (B) were infected with lentiviruses expressing indicated mCherry-MTMR, fixed with 4% PFA, and imaged by confocal microscopy. Scale bar—10 μm. Asterisks indicate the restored PM localization of GFP-KRASG12V or GFP-LactC2. (C–F) Intact basal PM sheets were prepared from T47D cells co-expressing MTMR 4 shRNA66 and GFP-KRASG12V (C) or GFP-LactC2 (D), GFP-P4M-SidM (E), or GFP-2xFYVE (F), and infected with lentiviruses expressing indicated mCherry-MTMR members, and labeled with anti-GFP–conjugated gold particles and visualized by EM. The graphs show a mean number of gold particles ± SEM (n = 10). Significant differences between control (MTMR 4 KD and untagged mCherry-expressing) cells and other cells were assessed by one-way ANOVA tests for C–F (*P < 0.05, **P < 0.01, ***P < 0.001, #—not significant).