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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: miR-214-3p-Sufu-GLI1 is a novel regulatory axis controlling inflammatory smooth muscle cell differentiation from stem cells and neointimal hyperplasia

Fig. 1

miR-214-3p inhibits TNFα-induced inflammatory SMC differentiation from AdSPCs. ac Passage 3–8 AdSPCs were cultured in SMC differentiation medium in the absence (vehicle (Veh)) or presence of 50 ng/ml TNFα for 4 days to allow for inflammatory SMC (iSMC) differentiation. Total RNA, protein and conditioned culture medium were harvested and subjected to RT-qPCR (a), Western blot (b) and ELISA analyses (c), respectively. df miR-214-3p over-expression rescued mature/contractile SMC marker expressions and inhibited TNFα-induced inflammatory cytokine gene expression. AdSPCs were differentiated into iSMCs (DMEM containing 5 ng/ml TGFβ1 and 50 ng/ml TNFα) for 4 days, then transfected with miR-214-3p mimics (miR-214) or negative control (miR ctrl) and cultured for 48 h in the same culture medium. Total RNA, proteins and conditioned culture medium were harvested and subjected to RT-qPCR (d), Western blot (e) and ELISA (f) analysis, respectively. The data presented here are representative (top panels in b and e) or mean ± S.E.M. (a, c, d, f and bottom panels in b and e) of five or six independent experiments (n = 5 or 6). *P < 0.05 (vs vehicle/control); #P < 0.05 (TGFβ1/TNFα vs TGFβ1 or TNFα/miR-214 vs TNFα/miR ctrl) (one-way ANOVA with a post hoc test of Tukey’s analysis). miR-214 indicates miR-214-3p

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