Fig. 5: Negative regulation of PKCγ expression by ΔNp63α is dependent on miR-320a.

a A putative miR-320a binding site in PKCγ 3′UTR identified by Target Scan. b A431 cells were co-transfected with a luciferase reporter carrying PKCγ 3′UTR or a random 3′UTR along with control mimic or miR-320a mimic. At 24 h post transfection, a luciferase reporter assays were performed. y-Axis represents the relative change in the luciferase activity. Significant changes (P ≤ 0.05) relative to control mimic are indicated with an asterisk. c HaCaT cells were transfected with either NSC siRNA or p63 siRNA in conjunction with a negative control mimic or miR-320a mimic as indicated. Total RNA was extracted and transcript levels of ΔNp63α and PKCγ was measured by Taqman based qRT-PCR (upper panel). y-Axis represents the fold change in ΔNp63α and PKCγ transcript levels relative to NSC-transfected cells. Error bars indicate standard deviation. Significant changes (P ≤ 0.05) relative to respective NSC controls are indicated with an asterisk. The change in indicated protein levels were measured analyzed via immunoblotting with p63, PKCγ, Rac1, and pRac1 (S71) antibodies as indicated (lower panel). Immunoblot with β-actin was performed to confirm equivalent protein loading