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. 2021 Dec 23;10:e60311. doi: 10.7554/eLife.60311

Figure 6. HAT enzyme Kat2a was required for 8 C mediated histone acetylation to inhibit oxidative stress in heart protection.

(A) Western blot of H3K9ac level showed Kat2a knockdown reduced 8C-induced H3K9ac increase in NRVMs under both normoxia and sI/R. (B) Measurement of medium LDH levels in NRVMs at indicated condition using LDH assay kit. (C) FACS analysis of DHE staining NRVMs after sI/R. (D) Relative mean fluorescence intensity of DHE staining. (E) Enrichment of H3K9ac over H3 at promoters of HO1 and NQO1 after sI/R at indicated conditions. (F) mRNA expression of HO1,NQO1 and Kat2a in NRVMs after sI/R. (G) Schematic diagram of 8 C metabolism for cardiac repair. n = 3, *p < 0.05, **p < 0.01, ***p < 0.001, vs Normoxia + PBS + shCTL; #p < 0.05, ##p < 0.01, ###p < 0.001 vs sI/R + PBS + shCTL. Data were analyzed by two-way ANOVA, followed by post-hoc Tukey test.

Figure 6—source data 1. Original numeric data for Figure 6.
Figure 6—source data 2. Original western blot figure for Figure 6.

Figure 6.

Figure 6—figure supplement 1. Knockdown of Kat2a is required for alleviating ROS accumulation.

Figure 6—figure supplement 1.

(A) Kat2a was upregulated after I/R based on RNA-seq results. (B) Western blot showed knockdown of Kat2a by adenovirus sh-RNA. (C) Cell viability measurement in NRVMs at indicated condition using CCK8 detection kit. (D) NRVM cellular ROS levels are indicated by DHE staining after sI/R treatment. Scale bar: 200 µm. (E–G) Western blot and quantifications of HO1 and NQO1 in NRVMs after sI/R. n = 3, *p < 0.05, **p < 0.01, vs Normoxia + PBS + shCTL; # p < 0.05 vs sI/R + PBS + shCTL. Data were analyzed by two-way ANOVA, followed by post-hoc Tukey test.
Figure 6—figure supplement 1—source data 1. Original western blot figure for Figure 6—figure supplement 1.