Fig. 4

Reducing EP300 activities rebalances histone acetylation and rescues proliferation and differentiation deficits of FMRP-deficient NPCs. a FMRP protein expression in primary Fmr1 KO and WT NPCs treated with vehicle or curcumin (Cur). b A schematic model showing that reducing EP300 activities either pharmacologically or genetically rebalances histone acetylation and rescues proliferation and differentiation of Fmr1 KO NPCs. c–e Western blot analyses of total and acetylation of histone H3 (c, d) and histone H4 (c, e) in NPCs treated with curcumin (n = 3). Total histone H3 and total histone H4 were used as loading controls for acetylated H3 and H4, respectively. f, g Curcumin treatment rescued the cell proliferation phenotype of Fmr1 KO NPCs as assessed by incorporation of the thymidine analog, BrdU (f, red, BrdU; blue, DAPI. scale bar, 20 µm) and quantitative analysis (g, n = 3). h, i Curcumin treatment rescued neuronal differentiation phenotypes of Fmr1 KO NPCs as assessed by a neuronal marker Tuj1⁺ (h, red, Tuj1; blue, DAPI. scale bar, 20 µm) followed by quantitative analysis (i, n = 3). j, k Acute knockdown of EP300 rescued the cell proliferation phenotype of Fmr1 KO NPCs as assessed by incorporation of the thymidine analog, BrdU (j, red, BrdU; green, GFP, scale bar, 20 µm) and quantitative analysis (k, n = 3). l, m Acute knockdown of EP300 rescued neuronal differentiation phenotypes of Fmr1 KO NPCs as assessed by a neuronal marker Tuj1⁺ (l, red, Tuj1; green, GFP, scale bar, 20 µm) followed by quantitative analysis (m, n = 3). *P < 0.05, **P < 0.01, ***P < 0.001. n.s.: no significant difference. Two-way ANOVAs were used for data analyses. Data are presented as mean ± s.e.m.