Fig. 5

MITF binds to the ADAM10 promoter. a UCSC genome browser screenshot showing a biological replicate ChIP-seq of MITF bound to the ADAM10 locus [40]. b Histogram represents ChIP assays in IG37 cells (*p < 0.05). Agarose gels using the PCR products of the ChIP experiments on 501mel cell lines are presented. c HEK293 cells were transfected either with a plasmid containing the promoter sequence of ADAM10 upstream of the luciferase-coding sequence (Promoter WT) or with a plasmid contained an ADAM10-mutated promotor sequence (Promoter MUT) in the absence or the presence of overexpressed MITF. The analysis of MITF expression was carried out by western blot assays by using an anti-FLAG antibody, while actin protein was used as load control. *p < 0.05 when compared with MITF negative cells. **p < 0.05 when compared with promoter WT. NT, negative transfected cells. d Western blots of different melanoma cell lines, both MITF positive (501mel, SK-MEL-28 and IGR37) and MITF negative (A375, SK-MEL-2, and IGR39) to analyze the levels of MITF and ADAM10 proteins. e Western blots to analyze N-Cad, MITF, and ADAM10 proteins in IGR37 cells (MITF positive) and IGR39 cells (MITF negative). f Western blots to analyze N-Cad, MITF, and ADAM10 proteins in IGR37 cells and several MITF-KO clones. g Effect of MITF silencing on the expression of N-Cad, MITF, and ADAM10 in SK-MEL-28 cells. Blots in this figure were cropped from different gels. Full blots are shown in Fig. S9 and quantified in Fig. S8