PPP6C expression affects MITF expression and drug resistance in melanoma. (A) Real-time PCR measurement of PPP6C in cells 72 h after transfection with either siPPP6C or siNon-targeting (Student’s two-tailed t-test). (B) Real-time PCR measurement of MITF and three target genes in cells with and without PPP6C knockdown 72 h after transfection (Student’s two-tailed t-test). (C) Drug dose response curve for A375 cells treated with dabrafenib and knockdown of PPP6C 72 h after transfection and treatment. Dotted lines display EC 50 for each condition. (D) Cellular expression of total MITF in A375 cells. Representative confocal images of fluorescence 72 h after transfection with either siPPP6C or siNon-targeting and staining with a MITF antibody, with quantification of fluorescence for each condition. One of two independent experiments is shown (Student’s two-tailed t-test). (E) Cellular expression of phospho-MITF in A375 cells. Representative confocal images of fluorescence 72 h after transfection with either siPPP6C or siNon-targeting and staining with a phospho-specific MITF antibody, with quantification of fluorescence for each condition. One of two independent experiments is shown (Student’s two-tailed t-test). p-values are indicated as following: *p < 0.05, **p ≤ 0.01. NS: non-significant p-value.
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