Fig. 5.

AT₁R is a novel target of miRNA let-7b. a: PASMCs were transfected with 50 nM miCon, let-7b-mimic, iCon or iLet-7b for 48 h. AT₁R mRNA was detected by RT-qPCR. b: PASMCs were transfected with 50 nM miCon or let-7b-mimic at a concentration of 10 nM, 50 nM, 100 nM, 150 nM or 200 nM for 48 h. c: PASMCs were transfected with 50 nM iCon or iLet-7b at a concentration of 10 nM, 50 nM, 100 nM, 150 nM or 200 nM for 48 h. AT₁R protein was detected by Western Blot. d: The diagram shows the structure of AT₁R 3’UTR luciferase in pGL3 reporter. The binding sites of let-7b in the AT₁R 3’UTR sequence and the corresponding mutant sites (shown in green). e: Dual luciferase reporter assay. HEK-293 T cells were cotransfected with 200 ng of pGL3-AT₁R-3’UTR-WT or pGL3-AT₁R-3’UTR-Mut and 20 ng of Renilla plasmid. miCon or let-7b-mimic at a concentration of 10 nM, 20 nM and 40 nM was also added. Values were presented as means ± SD from 3 independent experiments. **0.001 ≤ P ≤ 0.009 (different from the corresponding control); ***P < 0.001 (different from the corresponding control)