Figure 5.

βHB may upregulate Trx1 through HDAC inhibition. (A) Dosage effects of βHB on protein acetylation and HDAC1 expression. Cardiomyocytes were treated with the indicated concentrations of βHB for 16 h. n = 4–5. (B) Dosage effects of H₂O₂ upon HDAC1 expression. Cardiomyocytes were treated with the indicated concentrations of H₂O₂ for 16 h. n = 3–4. (C) Knockdown of HDAC1 upregulates Trx1. (Left) Cardiomyocytes were transduced with 0, 0.3, 1, 3, 10, or 30 MOI of shHDAC1 for 3 days. (Right) Trx1 induction by shHDAC1 (3 MOI). n = 7–9. (D) βHB induces Trx1 acetylation. Flag- and HA-tagged Trx1 was expressed with adenovirus vector and immunoprecipitated with anti-Flag antibody. n = 4–6. (E) βHB inhibits Trx1 degradation. Cardiomyocytes were treated with 100 µM cycloheximide (CHX) for 1 h followed by 1 mM βHB for 1 to 4 h. n = 4–5. (A–E) Western blot analyses were performed with indicated antibodies. * p < 0.05 (A–E).