Figure 5.

HCAL Regulates LAPTM4B Expression by Competitively Binding to Common miRNAs

(A) Schematic representation of an RNA immunoprecipitation assay. (B) MS2-RIP followed by miRNA qPCR to detect endogenous miRNAs associated with HCAL. (C) Relative luciferase activity in SMMC-7721 cells cotransfected with the indicated miRNAs and luciferase reporter vectors containing HCAL. (D) SMMC-7721 cells were transfected with the indicated miRNAs. After 48 hr, relative expression of HCAL was analyzed by qPCR. (E) RIP assay using anti-AGO2 antibody and qPCR were performed with SMMC-7721 cells transiently overexpressing the indicated miRNAs to detect the association of HCAL with AGO2. (F) Changes in LAPTM4B mRNA expression levels in the indicated SMMC-7721 cells. (G) Changes in the luciferase activity of LAPTM4B 3′ UTR in the indicated SMMC-7721 cells. Data are presented as the relative ratio of firefly luciferase activity to Renilla luciferase activity. Data are expressed as mean ± SD. *p < 0.05 and **p < 0.01.