PDK1 regulates MRCKα localization in lamellipodia. (A–C) Evaluation of endogenous MRCKα localization in lamellipodia. In MCF10A cells, stained by immunofluorescence against MRCKα, lamellipodia (identified with F-actin) were divided in three classes on the level of MRCKα accumulation. MCF10A cells were tested for MRCKα accumulation in the presence or absence of EGF stimulation or, in the case of stable transduction, with: shScr, shPDK1 #79, shPDK1 #81, PDK1_WT, PDK1_KD, PDK1_ΔPH, PDK1_L155E, PDK1_Δ50, or empty lentiviral vectors. A number equal or superior to 100 cells was counted each condition. Each error bar represents the mean and standard error of the mean of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Bars, 10 µm.