Research Paper Volume 15, Issue 8 pp 2877—2890

Figure 6. MAPK1 was a direct target of miR-324-3p in CMM progression. (A) Kyoto Encyclopedia of Genes and Genomes analysis revealed that the MAPK signaling pathway was significantly altered in CMM. (B) The effect of LINC01296 on MAPK1 expression was analyzed by Western blotting with the indicated antibodies and samples from the M21 and A2058 cells transfected with sh-LINC01296 or sh-NC. (C) The relative expression of MAPK1 in M21 and A2058 cells transfected with knocking down LINC01296 by qRT-PCR analysis. (D) Schematic illustration of the predicted binding sites between miR-324-3p and MAPK1, and mutation of potential miR-324-3p binding sequence in MAPK1. Relative luciferase activities of wild type (WT) and mutated (MUT) MAPK1 reporter plasmid in human embryonic kidney (HEK) 293T cells co-transfected with miR-324-3p mimic. (E) MAPK1 expression level in CMM cell lines and human epidermal melanocytes, adult cell line (HEMa-LP) were detected by qRT-PCR analysis. (F) The relative expression of MAPK1 in M21 and A2058 cells transfected with miR-136-5p mimic by Western Blotting analysis. (G) The relative expression of MAPK1 in M21 and A2058 cells transfected with miR-136-5p mimic by qRT-PCR analysis. All of data were analyzed from three independent experiments. *P < 0.05, ** P < 0.01, ^ns P>0.05.