Fig. 6.

Stable expression of miR-211–5p reduces sensitivity to vemurafenib. (A) Representative images of the MML-1 cells transfected with scrambled (scr) or miR-211 vectors. (B) The relative expression of miR-211–5p in MML-1 cells transfected with the miR-211–expressing lentiviral vector compared with MML-1 cells transfected with the scrambled gene-expressing vector. C. Elegans miR-39–3p was used as the external spike-in control to normalize the expression. (C) Cell counting shows that miR-211–5p–transfected cells proliferate more than the cells transfected with the scrambled gene. Cells were counted with the trypan blue exclusion assay. (D) Fold change in miR-211–5p expression with respect to C. elegans miR-39–3p between the scrambled and miR-211–transfected cells upon vemurafenib treatment. (E) Relative percent cell proliferation measured by MTT assay. The MTT assay was performed after 72 h of vemurafenib treatment. (F) Fold change regulation of miR-211–5p in MML-1R cells compared with the sensitive parental MML-1 cells. The fold change was normalized to the external spiked-in C. elegans miR-39–3p. (G) MML-1R cells were plated onto a 96 well plate at a density of 10,000 cells per well. Cells were transfected with control oligos or miR-211–5p inhibitors and the cellular proliferation was assessed by MTT after 24 h by using absorbance spectrum at 570 nm. All experiments were performed three times independently. Data are presented as ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (Fig. 5 legend for repeated abbreviations.) MML-1R, MML-1 resistant cells; MML-1S, MML-1 sensitive cells.