Research Paper Volume 8, Issue 2 pp 272—290

Figure 3. miR-603 directly targets LRPAP1 by binding the 3′UTR of LRPAP1 and downregulates LRPAP1 mRNA and protein levels. (A) Alignment of potential binding sites for miR-603 in the 3′UTR of LRPAP1 by PITA; scrambled sequences are also included. (B) Verification of the miR-603 target sites in the 3′UTR of LRPAP1 by dual-luciferase reporter assay. (C) Quantitative RT-PCR analysis of mRNA (right) and immunoblot analysis of protein (left) levels of LRPAP1 after the overexpression of miR-603 in HEK293 cells. (D) Quantitative RT-PCR analysis of mRNA (right) and immunoblot analysis of protein (left) levels of LRPAP1 following the inhibition of endogenous miR-603 in HEK293 cells. Quantitative RT-PCR analysis of miR-603 following its inhibition (right) in HEK293 cells. (E, F) Quantitative RT-PCR analysis of mRNA (right) and immunoblot analysis of protein (left) levels of LRPAP1 following the overexpression of miR-603 (E) and the inhibition of endogenous miR-603 (F) in HeLa cells. *P < 0.05, **P < 0.01, ***P < 0.001 versus NC as determine by paired two-tailed Student's t-tests for luciferase activity analysis, quantitative RT-PCR analysis (B-F), and immunoblot analysis (B-D) and by unpaired two-tailed Student's t-tests for immunoblot analysis (E, F).