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. 2015 Jan;185(1):252–265. doi: 10.1016/j.ajpath.2014.09.012

Figure 5.

Figure 5

MITF regulates PEDF gene expression in vivo, as demonstrated by both DN and siRNA gene silencing methods. A and B: qPCR analysis of PEDF mRNA levels in RPE cells (ARPE19) (A) and primary melanocytes (B) after infection with adenovirus expressing a random polypeptide control, WT MITF, or DN MITF shows a marked increase in endogenous PEDF mRNA levels by functional MITF and a decrease by nonfunctional MITF in RPE cells and melanocytes, compared with control. C: In accord, a marked increase in PEDF protein levels was demonstrated in melanocytes expressing WT MITF, and a marked decrease in DN MITF melanocytes, compared with control. D: Similarly, marked increase and decrease in PEDF mRNA levels were demonstrated in melanoma cell line (UACC62) expressing WT and DN MITF, respectively, compared with control. E and F: In two independent experiments, siMITF markedly decreased both MITF and PEDF mRNA levels in melanocytes (E) and in the melanoma cell line SK-MEL-5 (F). Ctrl, control; DN, dominant negative; WT, wild type.