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. 2017 Aug 10;108(9):1811–1819. doi: 10.1111/cas.13315

Figure 3.

Figure 3

MiR‐372 directly targets ULK1 and regulates autophagy. (a) Detection of dual luciferase reporter gene activity: BXPC‐3 cells were co‐transfected with miR‐372 mimics and ULK1 3′UTR WT/Mut recombinant plasmids. The luciferase activity assay suggested that miR‐372 inhibits the luciferase activity of WT plasmid but had no effect on the activity of Mut plasmid. (b) ULK1 expression on mRNA level as determined by qRT‐PCR in BXPC‐3 and PANC‐1 cells transfected with miR‐372 mimics and anti‐miR‐372 or control. (c) Total protein and phosphorylation levels of ULK1 were determined by miR‐372 mimics and anti‐miR‐372 transfected cells by western blotting in BXPC‐3 and PANC‐1 cells. (d) P62 and LC3 levels were determined in miR‐372 mimics and anti‐miR‐372 transfected cells by western blotting. GAPDH was used as an internal control. (e) Immunohistochemical analysis of ULK1 expression in pancreatic adenocarcinoma tissues and normal pancreas tissues. *P < 0.01 versus negative control (NC).