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. 2017 Jan 17;14(3):326–338. doi: 10.1080/15476286.2016.1276149

Figure 4.

Figure 4.

MiR-148a targets PPARGC1A, and miR-17–5p targets PPARA (A) PPARGC1A expression level is quantified by RT-qPCR (n = 6) GMECs are transfected with miR-148a mimic or inhibitor for 48h, and. White bars: negative control; black bars: miR-148a mimic or inhibitor. (B and C) Target site of miR-148a in the PPARGC1A 3′-UTR and the construction of the luciferase (Luc) expression vector fused with the PPARGC1A 3′-UTR. WT: Luc reporter vector with the WT PPARGC1A 3′-UTR (1085 to 1106); MU: Luc reporter vector with the mutation at the miR-148a site in PPARGC1A 3′-UTR. (D) Western blot analysis of PPARGC1A expression in the miR-148a mimic and NC treatment experiments. The effect of miR-148a mimics and Inhibitor on PPARGC1A protein expression in GMECs was evaluated by western blot analysis. Total protein was harvested after 48 h post-transfection, respectively. (E) PPARA expression quantified by RT-qPCR (n = 6) in GMECs transfected with miR-17–5p mimic or inhibitor for 48 h. White bars: negative control; black bars: miR-17–5p mimic or inhibitor. (F and G) Target site of miR-17–5p in PPARA3′-UTR and the construction of the luciferase (Luc) expression vector fused with the PPARA3′-UTR. WT: Luc reporter vector with the WT PPARA 3′-UTR (419 to 425); MU: Luc reporter vector with the mutation at miR-17–5p site in PPARA 3′-UTR. (H) Western blot analysis of PPARA expression in the miR-17–5p mimic and NC treatment experiments. The effect of miR-17–5p mimics and Inhibitor on PPARA protein expression in GMECs was evaluated by Western blot analysis. Total protein was harvested after 48 h post-transfection. All experiments were duplicated and repeated 3 times. Values are presented as means ± standard errors, *, P < 0.05; **, P < 0.01.