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. Author manuscript; available in PMC: 2023 Jun 15.
Published in final edited form as: Circulation. 2022 May 18;145(24):1784–1798. doi: 10.1161/CIRCULATIONAHA.121.057868

Figure 7. Kynurenine activates assembly of RUNX2 and CUL4BAhR E3 ligase complexes.

Figure 7.

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.