SMURF2 Antibody - #DF7683

Fig 6 The YBX1/CBX3 axis promotes tumor growth via suppressing SMURF2 in pancreatic cancer. (A-D) PANC-1 and SW1990 cells were transfected with indicated shRNAs for 72 hours. After puromycin selection, cells were harvested for the RT-qPCR (A), MTS (B), and colony formation (C and D). (E-G) PANC-1 cells were transfected with indicated shRNAs for 72 hours. After puromycin selection, cells were subcutaneously injected into the nude mice. The image of tumor was shown in panel E. The tumor mass and tumor growth curve were demonstrated in panel F and G, respectively. Data are presented as mean ± SD (n = 5). (H) PANC-1 and BxPC-3 cells were transfected with indicated shRNAs for 72 hours. After puromycin selection, cells were harvested for the Western blot analysis. (I) PANC-1 and BxPC-3 cells were transfected with indicated shRNAs for 48 hours, and then were transfected with indicated constructs for another 24 hours. Cells were harvested for Western blot. (J) A hypothetic model depicted that CSE exposure-induced YBX1 overexpression contributed to the upregulation of CBX3, which inhibited SMURF2 expression to promote the progression of pancreatic cancer. Data in (A), (B) and (D) are presented as mean ± SD (n = 3). Not significant Ns; P < 0.001 ***.

Figure 7 TGF-β2 induces expression of TFPI2 in vitro. Human renal glomerular endothelial cells (hRGECs) were incubated with human recombinant protein TGF-β2 to induce EndMT in vitro. A, the expression of TFPI2 and SMURF2 in hRGECs following treatment of 0, 1, 2.5, 5, or 10 ng/ml TGF-β2 for 48 h. B, the expression of TFPI2 and SMURF2 in hRGECs following treatment of 5 ng/ml TGF-β2 for 0, 12, 24, or 48 h. To explore the molecular mechanism of TGF-β2-induced TFPI2 expression, we next verified whether TFPI2 is a direct target gene of TGF-β/Smad signal. According to JASPAR prediction, TFPI2 is a potential target gene of SMAD2/3/4. C, the promoter activity of TFPI2 gene under TGF-β2 stimulation was detected by Dual-Luciferase Reporter Assay. D, SMAD4 was knocked down in hRGECs in the presence of TGF-β2, which was confirmed by Western blot. E, the expression of TFPI2 was detected in SMAD4-silencing cells. Data are shown as the mean ± SD (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. hRGEC, human renal glomerular endothelial cell; SMURF2, SMAD ubiquitination regulatory factor-2; TFP12, tissue factor pathway inhibitor 2; TGF-β, transforming growth factor beta.