Figure 3.

Deacetylation contributes carbons to AcCoA and downstream pathways.A, schematic of histone deacetylation contributing to AcCoA. B–F, ¹³C enrichment during pulse-chase experiment. N > 9 across at least three independent experiments. G–I, AcCoA labeling during pulse-chase experiment with ACSS2i (22 μM). G, ¹³C enrichment (H) raw signal intensity (area under the curve (AUC)) for indicated isotopologues. I, ¹³C enrichment after washout with washout enrichment subtracted. J–L, same as (G–I) but for (iso)butyryl-CoA. N = 9 across three independent experiments. M–O, quantitation of cellular acyl-CoAs with the portion derived from protein-bound acetate indicated in red (calculated by multiplying percent molar enrichment by pool size at each time point). All data indicated as mean ± SD. Statistical analyses in (B–D) was by two-way ANOVA followed by Sidak’s multiple comparison test, and I and L were by two-way ANOVA followed by Dunnett’s Multiple Comparison. ns = not significant. p value < 0.05 (∗), p value < 0.01 (∗∗), p value < 0.001 (∗∗∗), and p value < 0.0001 (∗∗∗∗). A, created with BioRender.com. AcCoA, acetyl-CoA; ACSS2i, ACSS2 inhibitor.