Figure 5

Figure 5

(a–d) Gene expression of ECM and EMT genes evaluated by RT-qPCR in EV-SkMel28 and MITF-KO cells with ectopic expression of EV-FLAG and MITF-FLAG constructs. Expression was normalized to EV-SkMel28 cells. Error bars represent standard error of the mean, * p value <0.05 was calculated by two-way ANOVA (multiple correction with Sidak test). (e) Schematic showing the dox-inducible MITF KD system. MITF expression decreases in the presence of dox (first 24 hr) and reverts back to baseline levels upon dox wash-off (at 72–96 hr). (f) Western blot analysis for the protein expression of MITF and CDH1, and CDH2 with the presence of dox treatment 0 and 24 hr or absence of dox 72 and 96 hr. (g–j) RT-qPCR analysis of MITF targeted ECM genes in miR-NTC and miR-MITF SkMel28 cells, treated with dox for 24 hr to induce MITF KD and after dox wash-off at 72 and 96 hr. Expression was normalised to miR-NTC cell lines, error bars represent standard error of the mean, * p value <0.05 was calculated by two-way ANOVA (multiple correction with Sidak test).