Fig. 2.

Sonic hedgehog-glioma-associated oncogene 1 (Shh-GLI1) signal activation and modulation by miR-214-3p during iSMC differentiation. a, b Luciferase activity assays to examine Shh-GLI signalling activity. AdSPCs were transfected with GLI reporter constructs and subjected to various treatments indicated in respective figures. Two days later, total cell lysates were harvested and subjected to luciferase activity assays. c, d RT-qPCR detection of Shh-GLI target gene expression. e miR-214-3p promotes GLI1 nuclear translocation. AdSPCs were subjected to similar treatments described in Fig. 1d–f. Total cell lysate and nuclear proteins were extracted and subjected to Western blot analysis. The data presented here are representative (left panel in e) or mean ± S.E.M. (a–d and right panel in e) of five independent experiments (n = 5). *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