Fig. 7

SENP1 regulates LR-MSC myofibroblast transition through both the WNT/β-Catenin and Hedgehog/GLI signaling pathways in vitro. A mRNA expression of endogenous SENP1, β-Catenin, and GLI1 were assessed using RT-PCR in different groups. RT-PCR analysis indicating that β-Catenin and GLI1 mRNA levels did not change in the presence of LV-SENP1-shRNA during TGFβ1 induction. *P < 0.01, **P < 0.05, ^▲P < 0.01, ^▲▲P > 0.05. *, the control group vs. the TGFβ1 group; **, the SENP1 knockdown group vs. the TGFβ1 group; ^▲, the control group vs. the TGFβ1 group; ^▲▲, the SENP1 knockdown group vs. the TGFβ1 group. B SENP1 level in total cell lysates was assessed using western blot analyses. *P < 0.01, **P < 0.05. *, the control group vs. the TGFβ1 group; **, the SENP1 knockdown group vs. the TGFβ1 group. C β-Catenin and GLI1 levels in total cell lysates were assessed using western blot analyses. The SUMOylation analysis of the immunoprecipitated β-Catenin and GLI1 proteins. β-Catenin and GLI1 were immunoprecipitated from whole-cell lysates using anti-β-Catenin and anti-GLI1 antibodies, respectively. The blots were probed with SUMO1. *P < 0.01, **P < 0.05, ^▲P < 0.01, ^▲▲P < 0.05. *, the control group vs. the TGFβ1 group; **, the SENP1 knockdown group vs. the TGFβ1 group (β-Catenin and GLI1); ^▲, the control group vs. the TGFβ1 group; ^▲▲, the SENP1 knockdown group vs. the TGFβ1 group (β-Catenin-SUMO and GLI1-SUMO). All data are presented as the mean ± SEM of three independent experiments. Quantification is shown in the lower panel. Each experiment was performed in triplicate.