ICAT negatively regulates the M-MITF promoter activity by competing with LEF1.
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Abstract
<p><b>A.</b> Mel501 cells were transfected with a <i>M-MITF</i>::<i>luciferase</i> vector in the presence of increasing amounts of <i>CMV</i>::<i>ICAT-WT</i> expression vector. Data are presented as means ± SEM of three independent experiments. <b>B.</b> qRT-PCR analysis of <i>M-MITF</i> mRNA levels in Mel501 cells transfected with empty or <i>ICAT-WT</i> expression vectors. <b>C.</b> WB analysis of MITF and ICAT proteins in Mel501 cells transfected with empty or <i>ICAT-WT</i> expression vectors. <b>β</b>-actin = loading control. <b>D.</b> WB analysis of MITF and p27<sup>Kip1</sup> protein levels in siRNA and ICAT-transfected Mel501 cells. SiMITF treatment and ICAT overexpression induce respectively a 42% and 35% increase of p27 protein amount; Scrb = control scrambled siRNA. <b>E</b>. Mel501 cells were transfected with <i>M-MITF</i>::<i>luciferase</i>, <i>LEF1</i> and <i>ICAT-WT</i> expression vectors. Data are presented as means ± SEM of three independent experiments. *p<0.05, **p<0.01, ***p< 0.001; ****p<0.0001.</p