Fig. 2

Treatment with sodium butyrate increases histone acetylation and affects ataxin-3 aggregation in SCA3 SH-SY5Y cells. (A) Western blots of SH-SY5Y stably expressing ataxin-3-84Q cells treated with 3 mM SB or vehicle control were probed for acetylated histone 3 at lysine 9 (ac-H3K9) and acetylated histone 4 at lysine 5 (ac-H4K5). (B) Densitometry of ac-H3K9 showed an increase with SB treatment compared to vehicle treatment (p = .0312). (C) Densitometry of ac-H4K5 revealed an increase with SB treatment compared to vehicle control (p = .0022, n = 6). (D) Immunostaining for ataxin-3 revealed a reduction in the amount of ataxin-3 positive aggregates after treatment with SB, compared to vehicle treatment (white arrows, scale bar: 10 μm). (E) Quantification of the number of ataxin-3 positive aggregates revealed decreased aggregates in SB-treated ataxin-3-84Q cells compared to the vehicle-treated control (p = .0022, n = 6). (F) Western blot of SH-SY5Y stably expressing ataxin-3-84Q cells treated with 3 mM SB or vehicle were probed for ataxin-3. (G) Densitometry of high molecular weight (HMW) ataxin-3 indicated that decreased HMW ataxin-3 was present following treatment with SB compared to vehicle control (p = .0156, n = 7). (H) Quantification of the ataxin-3 monomer levels following SB treatment increased compared to the vehicle control (p = .0156, n = 7). Data represent mean ± SEM. The statistical analysis was performed by Wilcoxon test. *p < 0.05, **p < 0.01.