Figure 4.

GSDMB-STAT3 signaling regulates IGFBP3 expression in bladder cancer. A-F. T24 and 5637 cell lines were infected with constructed plasmids (shGSDMB#1, shGSDMB#2, shSTAT3#1, shSTAT3#2 and Flag-GSDMB). After infecting 48h and 72h, all cells were harvested for RT-qPCR (B, D, F) and Western Blotting analysis (A, C, E). All data were showed as Means ± SD (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001. G. The schematic diagram of STAT3 binding to the promoter position of IGFBP3. H. T24 and 5637 cell lines were treated according to the protocol of ChIP experiment, and the DNA samples were performed DNA agarose gel electrophoresis. All data were showed as Means ± SD (n = 3). ***, P < 0.001. I-L. T24 cell lines were infected with constructed plasmids (shGSDMB, shSTAT3 and Flag-GSDMB). After infecting 48h and 72h, all cells were harvested for RT-qPCR (J and L) and Western Blotting analysis (I and K). All data were showed as Means ± SD (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, no significant. M-N. T24 cell lines were infected with constructed plasmids (shGSDMB and shSTAT3). Cells were harvested for MTS assay (M) and clone formation assay (N). All data were showed as Means ± SD (n = 3). *, P < 0.05; ***, P < 0.001; ns, no significant. O-Q. T24 cells were infected with constructed lentivirus to establish the stable cell lines. Then cells were injected subcutaneously into the nude mice to construct xenograft transplantation model. The image of xenografts was shown in (O), the tumor mass and volume were measured in (P and Q). All data were presented as Means ± SD (n = 6). ***, P < 0.001; ns, no significant.