Figure 5:

Inhibition of mIR-30c-2* and miR-497–5p targets affects cuSCC cell proliferation and survival. (A) SYBR green qRT-PCR of the validated miR-30c-2* and miR-497 targets in NHEKs and the cuSCC cell lines COLO16, SRB12, SRB1, IC1, and RDEB2. (B) Representative results of Western blotting of the validated miR-30c-2* and miR-497 targets in the same cell lines as in (A). (C) Western blotting of the indicated targets following siRNA-mediated knockdown in COLO16 cells. (D-F) COLO16 cells stably expressing nuclear mCherry transfected with the indicated siRNAs, incubated with Annexin V-488, and scanned every 4 hours using the Incucyte® high-content live-cell imaging platform. (D) Growth curve of COLO16 cells transfected with the indicated siRNAs. (E and F) Immunofluorescence (E) and quantification (F) for annexin V-488 (green)-positive cells transfected with the indicated siRNAs. (G) Cell cycle profiles of COLO16 cells were assessed by FACS 48 hours after siRNA transfection. Data shown are mean ± SD, of at least 3 independent experiments. * p<0.05, ** p<0.01, ***p<0.001, Student’s t test (two-tailed).