Figure 7

MiR-377 targets MAP3K7. (A) MAP3K7 mRNA levels in different melanoma cell lines relative to NHEM as assessed by qRT-PCR. (B) MAP3K7 protein levels in different melanoma cell lines as assessed by WB (left). The bar graph denotes the densitometry scanning of the total MAP3K7 bands normalized to the GAPDH bands (right). Data is represented as mean ± SEM of three independent experiments. (C) The 3′UTR of the MAP3K7 gene showing potential binding sites for miR-377 (marked), and the sequence conservation from human to lizard in the vicinity of miR-377 seed sequences (taken from TargetScan at http://www.targetscan.org/). (D) 293 T cells were co-transfected with a control empty psiCHEK-II-luciferase (vector), or wild type MAP3K7 3′UTR-luciferase plasmid, or mutant MAP3K7 3′UTR-luciferase plasmid, together with a plasmid expressing miR-377. The results are presented as the ratio of expression of renilla/luciferase that was normalized relative to control-transfected cells for each vector. Data is represented as mean ± SEM from three independent experiments. *signifies p < 0.05. (E) MAP3K7 mRNA levels in mel-14PA over-expressing HTR control vector or vector expressing miR-377 as assessed by qRT-PCR. (F) MAP3K7 protein levels in mel-14PA over-expressing HTR control vector or vector expressing miR-377 as assessed by WB.