Fig. 4: Nuclear Pdha1 promotes histone acetylation by increasing the local acetyl-CoA content.

a Relative content of acetyl-CoA in whole-cell, cytoplasm, or nucleus of MEFs during somatic cell reprogramming with SKOM. Cells after overexpression of Flag, nls-Pdha1, or nls-Pdha1 with triple mutation of its catalytic domain (Triple mutant) were tested. b, c Representative images (b) and quantification (c) of H3K9ac, H3K27ac, and H3ac (green) in MEFs with or without nls-Pdha1 overexpression. Scale bars, 5 µm. d Western blot analysis of H3K9ac, H3K27ac, and H3ac in MEFs with or without nls-Pdha1 overexpression, as well the band quantification. Anti-Flag was targeting nls-Pdha1 and Anti-H3 was used as a loading control. e H3K9me3, H3K27me3, and H3k36me3 modifications in MEFs with or without nls-Pdha1 overexpression were detected by western blot. Anti-Flag was targeting nls-Pdha1 and Anti-H3 was used as a loading control. The relative expression levels were quantified in the right panel. f, g Representative images (f) and quantification (g) of H3K9ac, H3K27ac, and H3ac (green) at day 4 of cell reprogramming with SKOM plus nls-Pdha1 or Flag (control). Scale bars, 5 µm. h The H3K9ac, H3K27ac, and H3ac modifications during cell reprogramming with or without nls-Pdha1 at indicated time point were detected by western blot. Anti-Flag was targeting nls-Pdha1 and Anti-H3 was used as a loading control. i The relative expression levels of H3K9ac, H3K27ac, and H3ac modifications during cell reprogramming with or without nls-Pdha1 at each time point were quantified. Data are presented as the mean ± S.D and n = 3 independent experiments (a, c–e, g, h, i). At least 30 cells were counted in each experiment (c, g). A two-tailed unpaired Student’s t test was used (a, c–e, g, i).