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. 2023 Jan 13;14(1):26. doi: 10.1038/s41419-022-05534-w

Fig. 7. Upregulation of Twist1 by KLF16 mediates the promoting effect of SF3B4 on ccRCC progression.

Fig. 7

A–C Caki-1 and 769-P cells were transfected with indicated constructs, and then western blot and RT-qPCR detected Twist1 expression. D The potential KLF16-binding motif on the proximal promoter of Twist1 gene. E ChIP-PCR was used to verify the binding site of KLF16 on the Twist1 promoter. F Dual-luciferase reporter gene examined the role of SF3B4 and KLF16 in regulating Twist1 promoter activity by co-transfection with indicated constructs. G, H Caki-1 cells were transfected with shTwist1 and oeKLF16, alone or together, and then transwell assay was used to examine cell migration and invasion. Scale bar = 50 μm. I The expression of E-cadherin and vimentin were detected by double immunofluorescence staining in cells transfected with indicated constructs. Scale bar = 20 μm. J, K Caki-1 cells were transfected with shSF3B4 and oeKLF16, alone or together, and then the expression of EMT-related genes was detected by western blot. L, M 769-P cells were transfected with oeSF3B4 and shTwist1, alone or together, and then the expression of EMT-related genes was detected by western blot. All data are expressed as the mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 vs. their corresponding controls.