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. 2016 Mar 22;7(18):26027–26041. doi: 10.18632/oncotarget.8266

Figure 4. MicroRNA-124-1 downregulates CASC3 expression by directly targeting its 3′-UTR.

Figure 4

A. Putative miR-124-1 binding sequence in the 3′-UTR of CASC3 mRNA. A human CASC3 3′-UTR fragment containing the wild-type or mutant miR-124-1 binding sequence was cloned downstream of the luciferase reporter gene. HEK293 cells were cotransfected with miR-124 or a control vector and a luciferase reporter construct containing the wild-type or mutant CASC3 3′-UTR. Luciferase activity was assayed 48 h after transfection. Firefly luciferase activity of each sample was normalized by Renilla luciferase activity. Data were normalized to the luciferase activity detected in cells transfected with the control vector, and the luciferase activity of the control vector was not significant. B. Effects of miR-124-1 overexpression on endogenous CASC3 expression as measured by real-time PCR and western blotting. β-actin served as the internal control. C. miR-124-1 and CASC3 protein expression were examined by in situ hybridization and immunohistochemistry, respectively.