A-C, Western blot analysis of human coronary VSMCs supplemented with DMSO or Kyn (50 μmol/L) followed by pretreatment with MG132 (1 μmol/L) before immunoprecipitation with anti-RUNX2 (A) or anti-AhR (B) or anti-CUL4B (C) antibodies and detection with indicated antibodies (n = 3).
D, Western blot analysis of RUNX2 expression in human coronary VSMCs transfected with Scr siRNA or AhR siRNA, followed by treatment with MG132 (1 μmol/L) for indicated periods (n = 4).
E, Western blot analysis of RUNX2 expression in human coronary VSMCs transfected with Scr siRNA or CUL4B siRNA, followed by treatment with MG132 (1 μmol/L) for indicated periods (n = 4).
F, Western blot analysis of RUNX2 expression in human coronary VSMCs transfected with RUNX2 plasmid combined with or without AhR and CUL4B plasmids, followed by CHX (50 μg/mL) treatment for the indicated periods (n = 4).
G, Western blot analysis of RUNX2 expression in human coronary VSMCs transfected with Scr siRNA or CUL4B siRNA, followed by treatment with DMSO or Kyn (50 μmol/L) (n = 4).
H, Western blot analysis of RUNX2 expression in human coronary VSMCs transfected with Vector or CUL4B plasmid combined with Scr siRNA or IDO1 siRNA (n = 4).
Results are presented as mean ± SEM, and analyzed using two-way ANOVA with Tukey’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, ns indicates P > 0.05.