Fig. 5. SMAD4 regulates FZD4 expression via SDNOR.

a FZD4 mRNA levels in porcine GCs after SDNOR knockdown, assessed by qRT-PCR. b Western blot analysis (left) and quantification (right) of FZD4, β-catenin protein levels in SDNOR-KD porcine GCs or in SMAD4^OE pGCs after SDNOR depletion, GAPDH used as control. c Western blotting analysis (top) and quantification (bottom) of FZD4 protein level in porcine GCs co-transfected siFZD4 with SDNOR^OE. d FACS analysis (left) and quantification (right) of the apoptosis rate of porcine GCs after FZD4 knockdown or with SDNOR overexpression. e Western blot analysis (left) and quantification (right) of c-Caspase-3 protein levels in porcine GCs after FZD4 knockdown or with SDNOR ectopic expression, GAPDH used as control. f FACS analysis (left) and quantification (right) of the apoptosis rate of porcine GCs after SMAD4 ectopic expression or with SDNOR knockdown. g Western blot analysis (left) and quantification (right) of c-Caspase-3 protein levels in porcine GCs after SMAD4 overexpression or with SDNOR knockdown, GAPDH used as control. Throughout, all data are shown as mean ± S.E.M. (n = 3, each). **p < 0.01 versus control or scrambled.