Fig. 5: Excess histones are hyperacetylated.
a Rad53-AID cells were adapted to normal or low glucose, Rad53 depletion was induced by addition of Auxin and tetracycline, and samples were collected at the indicated times to detect Rad53 and modified histones by western blot analysis (N = 3 replicates). The bar chart (right panel) provides a quantification of histone acetylation normalized by total histone levels. Significances were calculated with one-way ANOVA (pD_AcH3 = 9.4 × 10−5, pDR_AcH3 = 4.0 × 10−5, pDR_AcH4 = 7.4 × 10−5) with post hoc Tukey HSD test (pD_AcH3_2h_vs_UT = 0.26, pD_AcH3_4h_vs_UT = 0.0010, pD_AcH3_6h_vs_UT = 0.00013, pDR_AcH3_2h_vs_UT = 0.081, pDR_AcH3_4h_vs_UT = 0.00081, pDR_AcH3_6h_vs_UT = 0.000041, pDR_AcH4_2h_vs_UT = 0.12, pDR_AcH4_2h_vs_UT = 0.010, pDR_AcH4_2h_vs_UT = 0.000043). b Rad53-AID cells with the indicated SPT21 genotype were adapted to low glucose and treated as indicated with Auxin and tetracycline for 4h to deplete Rad53. Samples were collected to detect modified histones by western blot analysis. c Spt21-AID and Spt21S276A-AID cells were adapted to low glucose and Auxin for 16 h. Auxin was washed away to stabilize Spt21 and samples were collected after 6 h to quantify proteins by western blot analysis. Bars plots with error bars represent mean values and standard deviation, D glucose; DR glucose restriction (0.02% in liquid media); Aux Auxin; tc tetracycline. *p < 0.05, ***p < 0.001.
