Fig. 4

KLF4 upregulated NUCB2 expression in melanoma cells. a KLF4 WT or KO Mel-RM cells were treated with 3 μM TM at the indicated times. Gene expression profiles were obtained by RNA sequencing analysis. b The expression levels of MITF, VEGF, PDIA3, NUCB2 and IDH1 were analysed by q-RT-PCR. The data represent the means ± SD of three independent experiments; *p < 0.05, ***p < 0.001 vs. control. c The protein levels of KLF4 and NUCB2 were detected by western blot. d-e Flag-KLF4 and empty vector were individually transfected into KLF4 KO Mel-RM cells and the cells were treated with 3 μM TM at the indicated times. The expression levels of NUCB2 were analysed by q-RT-PCR and western blot assays. The data represent the means ± SD of three independent experiments; **p < 0.01 vs. control. f-g The Me1007 cells with or without KLF4 overexpression were treated using 3 μM TM at the indicated times. The expression levels of NUCB2 were analysed by q-RT-PCR and western blot assays. The data represent the means ± SD of three independent experiments; ***p < 0.001 vs. control. h-k Mel-RM and A375 cells with or without KLF4 knockdown were treated with 3 μM TM at the indicated times. The mRNA and protein levels of NUCB2 were detected by q-RT-PCR and western blot assays. Data represent the mean ± SD of three independent experiments. ***p < 0.001 vs. control