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. Author manuscript; available in PMC: 2011 Dec 10.
Published in final edited form as: Mol Cell. 2010 Nov 25;40(5):841–849. doi: 10.1016/j.molcel.2010.11.020

Figure 2.

Figure 2

Perturbing miR-211 but not melastatin affects melanoma invasion and motility. (A) Schematic presentation of the miR-211 gene in intron 6 of the melastatin (TRPM1) gene. (B) qRT-PCR of TRPM1 (black bars) normalized to actin, and mature miR-211 (gray bars) normalized to RNU48 in multiple human primary melanocytes (HP) and melanoma short term cultures (MSTCs). Y-axis is logarithmic scale. (N=5; means ± s.e.m.). (C) Matrigel assays for WM3682, WM3526, and 451LU (panels at left) transfected with control antagomir (Anti-miR-Ctrl, row 1), a miR-211-specific antagomir (Anti-miR-211, row 2), a control siRNA (siCtrl, row 3), or a melastatin-specific siRNA (siTRPM1, row 4). Membranes prior to scraping non-invasive cells from the top are shown (row 5). Matrigel assays for WM1745, WM1716, or WM3314 (panels at right) transfected with control miRNA mimic (Ctrl mimic, row 1) a miR-211 mimic (miR-211-mimic, row 2), a control Renilla cDNA (Renilla) or TRPM1 cDNA (TRPM1, row 3). The membranes prior to scraping cells from the top of the membrane are shown (row 4). Graphical presentation of the data quantifies the number of invasive cells in 10 high power fields (means ± s.e.m.; *p<0.005). (D) Modulating miR-211 but not melastatin affects melanoma motility. Real-time video microscopy of WM1716 (top panel) expressing Renilla cDNA, melastatin cDNA (TRPM1), a control miRNA mimic (Ctrl mimic), or miR-211 mimic. Real-time video microscopy of 451LU (bottom panel) expressing a control antagomir (Anti-miR-Ctrl), a miR-211 antagomir (Anti-miR-211) a siRNA control (siCtrl), or a melastatin siRNA (siTRPM1). Results quantify cell migration in μm/hr (>25 representative cells analyzed for each category from 2 movies, means ± s.e.m.; *p<0.005). Real-time video images are added as web-browsable links.