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. 2016 Aug 23;6:31506. doi: 10.1038/srep31506

Figure 3. miR-214 contributed to human MC hypertrophy by directly targeting PTEN in vitro.

Figure 3

(A) The mRNA levels of miR-214, PTEN. α-SMA. SM22 and Collagen IV. (B) The protein levels of PTEN, α-SMA, SM22 and Collagen IV. (C) Quantitative analysis of the protein levels of PTEN, α-SMA,SM22 and Collagen IV. LG + Osm, low glucose (5 mmol/L) supplemented with mannitol (20 mmol/L); HG, high glucose (25 mmol/L); ScRNA, HG with scramble control; anti-miR214, HG with transfection of miR-214 inhibitor. ▲p < 0.05 vs. HG, ★p < 0.05 vs. ScRNA. (D) A genome map for miR-214 binding sites of 3′ UTR of PTEN. (E) Schematic diagram indicating the sites of mutant of 3′ UTR of PTEN. (F) Dual luciferase measurement performed 48 hours after co-transfection of miR-214 mimics (MiR-214) with wild type (WT) or mutant (MU) of psi-CHECK-2-PTEN-3′UTR vector, respectively, in high glucose-stimulated MCs. ▲p < 0.05 vs. pGL4.10 Vector,★p < 0.05 vs. pGL4.10-WT Vector. (G) The mRNA levels of PTEN, α-SMA, SM22, Collagen IV and miR-214. (H) The protein levels of PTEN, α-SMA, SM22 and Collagen IV. (I) Quantitative analysis of the protein levels of PTEN, α-SMA, SM22 and Collagen IV. Control: MCs cultured in high glucose served as control; ScRNA: MCs transfected with scramble control; miR-214: MCs infected with lentiviral vectors expressing miR-214; miR-214+PTEN: MCs infected with lentiviral vectors expressing miR-214 and CDS of PTEN. si-PTEN: MCs transfected with siRNA for PTEN. The lentivirus-infected human MCs were cultured in high glucose for 48 hours. All experiments were performed in triplicate. All data were expressed as means ± SD, ▲p < 0.05 vs. control, ★p < 0.05 vs. miR-214+PTEN.