PKCε modulates RhoA in MK progenitors. (A) Western blot detection of PKCε, 14-3-3zeta, and RhoA protein expression in mouse MK cultured in the presence of TPO at indicated time points. GAPDH was monitored for protein loading. (B) Levels of PKCε, 14-3-3zeta, and RhoA protein expression during megakaryocytic differentiation of FLC. Densitometric measurements, performed using ImageJ software, of western blots from 3 replicates. Data are normalized for FLC (means ± 1 SD; *P < .05; **P < .01; ANOVA and Tukey tests). (C) Western blot detection of PKCε, 14-3-3zeta, and RhoA in mouse MK. At day 3 of culture, rMK untreated (un) or infected with PKCε-specific shRNA (shRNA59, shRNA60) and controls (shRNACT) were collected. GAPDH was monitored for protein loading. In the presence of PKCε-specific shRNA, the 14-3-3zeta protein levels remained stable at the control levels, whereas active RhoA expression levels sensibly increased. (D) Densitometric analysis of PKCε, 14-3-3zeta, and RhoA protein expression. Densitometric measurements, performed using ImageJ software, of western blots from 5 replicates. Data are normalized for shRNACT (means ± 1 SD; **P < .01 ANOVA and Tukey tests).