Fig. 5
From: miR-448 targets IDO1 and regulates CD8+ T cell response in human colon cancer
miR-448 downregulates IDO1 expression. a Luciferase reporter containing wild-type (WT) IDO1 3′ UTR was cotransfected with 40 miRNA mimics or a negative control (NC) into HCT-116 cells. Relative firefly luciferase expression was normalized to Renilla luciferase. NC: miRNA mimic negative control, 586: miRNA-586 mimic, etc. b The binding site of miR-448 in the IDO1 mRNA. The relative luciferase activity in c HT-29 cells and d HCT-116 cells cotransfected with miR-448 mimics and IDO1-WT, or cotransfected with miR-448 mimics and IDO1-MUT. e Representative western blots of IDO1 in HT-29 and HCT-116 cells transfected with miR-NC or miR-448 followed by IFN-γ treatment. GAPDH was served as the internal control. Images are representative of three biological replicates. f Representative ISH staining of miR-448 in human colon cancer tissues and adjacent noncancerous tissues. g IDO1 protein expression in miR-448low group (n = 29) and miR-448high group (n = 28). miR-448low group and miR-448high group were divided according to the ratio of miR-448 expression in colon cancer tissues to that in adjacent noncancerous tissues (cut-off = 1.33). a Mean ± SD; c, d, g mean ± SEM. c, d n = 9 measurements from three biological replicates performed in triplicate. a, c, d, g Two-tailed Student’s t-test was performed for statistical analysis; *P < 0.05, ***P < 0.001, NS: not significant. miR-NC: miRNA mimic negative control; miR-448: miRNA-448 mimic