Fig. 3
NRP2 expression depends on TGF-β/Smad signaling. a NRP2 protein expression after stimulation with 2.5 ng/mL TGF-β1 for 24 h as determined by Western blotting. Actin is shown as loading control. b qPCR analysis showing NRP2 mRNA levels in epithelial and mesenchymal-like HCC cell lines treated with either 2.5 ng/mL TGF-β1 or with 10 μM TGF-β inhibitor LY2109761 (LY) for 24 h. c SNU-449 hepatoma cells treated with control siRNA (siNT) and either administrated with 2.5 ng/mL TGF-β or 10 μM LY2109761 (LY) alone or in combination and analyzed for NRP2 and Smad4 expression by Western blotting. Smad4 knock-down (siSmad4) of SNU-449 cells treated with 2.5 ng/mL TGF-β1 for 24 h did not show modulation of NRP2 levels. Error bars depict SD from 3 independent experiments that were performed in triplicates