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. 2005 Jan 3;168(1):35–40. doi: 10.1083/jcb.200410115

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Copyright © 2005, The Rockefeller University Press

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Figure 3. — MITF regulates p16 ^Ink4a and Rb in normal melanocytes, but not in melanoma cells. (A) Real-time PCR assays of MITF and INK4A mRNA levels in 10T1/2 cells expressing ectopic MITF or an empty control vector (left panels) and in UM41 normal melanocytes transfected with MITF siRNA or control scrambled siRNA (right panels). Expression values represent fold difference compared with GAPDH. (B) Western blot analysis of MITF, p16^Ink4a, total Rb, Rb-phospho-Ser780, and tubulin protein expression in UM41 cells 48 h after transfection with plasmids expressing MITF siRNA or control scrambled siRNA. Arrows indicate hyperphosphorylated (top band) and hypophosphorylated (bottom band) forms of Rb. (C) BrdU incorporation assays in UM41 cells transfected with MITF siRNA or control scrambled siRNA. (D) Real-time PCR assays of MITF and p16^Ink4a mRNA levels in untransfected Mel202 melanoma cells. (E) Western blot analysis of MITF, p16^Ink4a, total Rb, Rb-phospho-Ser780, and tubulin protein expression in Mel202 cells 48 h after transfection with plasmids expressing MITF siRNA or control scrambled siRNA. (F) BrdU incorporation assay in Mel202 cells expressing MITF siRNA or control scrambled siRNA.