Fig. 7: ΔNp63α inhibits Rac1 phosphorylation and invasion by reducing PKCγ levels.

A431 cells were transfected with either NSC siRNA or siRNA targeting p63 for two rounds of transfections followed by treatment with DMSO or Gӧ6976 for 2 h as indicated. a The change in indicated protein levels were analyzed via immunoblotting with p63, Rac1, and pRac1 (S71) antibodies as indicated. Immunoblot with β-actin was performed to confirm equivalent protein loading. b The fold change in pRac1 levels relative to NSC DMSO-treated cells levels. Relative protein values are shown as means ± S.E.M. from n = 3 experiments. c At 24 h after the second round of transfection, 8.0 × 10⁴ cells were subjected to Matrigel-based invasion assay (c) and the number of invading cells was quantitated after 21 h. d The y-axis indicates the average number of cells invaded per field +1 standard deviation. Significant changes (P ≤ 0.05) relative to NSC controls are indicated with an asterisk. e A431 cells were incubated with DMSO or 100 nM of PMA for the indicated times. The change in indicated protein levels were analyzed via immunoblotting with Rac1 and pRac1 (S71) antibodies as indicated. f A431 cells were treated with DMSO or Gö6976 for 2 h and followed by incubation with 100 nM of PMA for 15 min. The change in indicated protein levels were analyzed via immunoblotting as indicated. g A431 cells were transfected with nonsilencing control siRNA (NSC) or siRNA specific to PKCγ followed by treatment with DMSO or 100 nM PMA for 15 min. Total RNA was extracted and transcript levels of PKCγ was analyzed by qRT-PCR. y-Axis represents the fold change in PKCγ transcript levels relative to NSC-transfected cells. The change in Rac1 and pRac1 levels was measured by immunoblot analysis (h). Immunoblot with β-actin was performed to confirm equivalent protein loading