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. 2015 Jun 29;43(16):7838–7849. doi: 10.1093/nar/gkv667

Figure 6.

Figure 6.

DDB2 modulates the TGF-β signaling via NEDD4L in human ovarian cancer cells. (A) Immunoblotting analysis shows that TGF-β-induced Smad2 phosphorylation is enhanced in DDB2-overexpressing CP70 cells. CP70 and three clones of DDB2-stably overexpressing CP70 cells were treated with TGF-β (5 ng/ml) for 4 h. Total cell lysates were prepared and subjected to immunoblotting analysis to determine the phosphorylation of Smad2. (B) qRT-PCR was conducted to analyze the TGF-β target genes. CP70 and CP70-DDB2 cells were treated with TGF-β for 4 h and total RNA was isolated for qRT-PCR analysis. (C) Concomitant knockdown of DDB2 and NEDD4L rescued the DDB2 downregulation-induced reduction of TGF-β signaling. DDB2 was knocked down alone or together with NEDD4L using their corresponding siRNA in PEO1 cells and treated with TGF-β for 4 h. Immunoblotting was carried out to detect the phosphorylation of Smad2. N = 3, Error bars represent SD. **P < 0.01 compared with non-TGF-β treated cells.