Fig. 6

JMJD6 is Transcriptionally Activated by c-Jun. a A375 cells were transfected with control siRNA or BRAF siRNAs. Total RNAs and proteins were extracted and analyzed for the expressions of BRAF and JMJD6 by qRT-PCR and Western blotting, respectively. b c-Jun recognized consensus site was identified in the promoter region of JMJD6 using a bioinformatics website (http://alggen.lsi.upc.es/cgi-bin/promo_v3/promo/promoinit.cgi?dirDB=TF_8.3). The number represents the nucleotide position relative to the transcription start site (+1). The mutated nucleotides are underlined. c Luciferase reporter assays in A375 cells transfected with JMJD6-Luc or mut-JMJD6-Luc together with c-Jun and renilla as indicated. d qChIP assays (upper) and ChIP assays (lower) of the occupancy of c-Jun in the promoter region of JMJD6 in A375 cells. FOXK1 and Igr5 intron 3 serve as positive and negative control, respectively. e A375 cells were transfected with vector, c-Jun, or treated with control siRNA or c-Jun siRNAs. Total RNAs and proteins were extracted and analyzed for the expression of JMJD6 by qRT-PCR and Western blotting, respectively. f Proposed model of the JMJD6 in melanoma carcinogenesis. In melanoma cancer cells, JMJD6, through regulating PAK1 alternative splicing and positively influencing the MAPK signaling, promotes melanogenesis, cell proliferation, invasion and angiogenesis. Hyperactive MAPK signaling leads to the phosphorylation of c-Jun, which, in turn, transcriptionally activates JMJD6 expression. Such a self-enhancing molecular system favors the development and progression of melanoma