FIGURE 5.

STAT4 is involved in the mechanism by which Occludin (OCLN) regulates IL8/STAT3 signalling to mediate bladder cancer angiogenesis. (A) Relative IL8 mRNA levels were detected in control and STAT4‐overexpressing 5637 and T24 cells using RT‐qPCR. (B) The STAT4 plasmid or the indicated vector was transfected into 5637 and T24 cells, and tube formation by EA.hy926 cells incubated with CM derived from the indicated cells was assessed using staining with Calcein AM and imaging with a fluorescence microscope. Scale bar = 100 µm. (C) The segment lengths were analysed, and the meshes were quantified using ImageJ software. (D) The relative STAT4 mRNA levels were detected in control and OCLN shRNA‐transfected 5637 and T24 cells. (E) Cells were transfected with the indicated siRNAs or plasmids for 24 h. The relative knockdown and overexpression efficiency was determined using Western blotting and RT‐qPCR. (F) 293T cells were cotransfected with the indicated plasmids for 24 h, and the relative activity of the IL8 promoter was evaluated by performing a dual luciferase reporter assay. The results are shown as the mean ± SD. *.01< p < .05; **.001< p < .01