Figure 1

HDAC3 stabilizes ataxin-7 and enhances post-translational modifications. (A) Ataxin-7-10Q (1–266) and ataxin-7-92Q (1–266) were transiently transfected with HDAC constructs [2–8] in HEK293T cells for 72 hrs. Lysates were collected and separated via SDS-PAGE on 4-12% bis-tris gels. Western blot transfer and analysis with an antibody to ataxin-7 (PA1-749) revealed the presence of bands of expected molecular weight for the ataxin-7 stop constructs as well as bands of higher molecular weight indicated by asterisks, particularly evident in the HDAC3 co-tranfection. GAPDH was used as a loading control. (B) Ataxin-7-10Q, ataxin-7-10Q (1–266), ataxin-7-10Q (1–266) K257R, ataxin-7-92Q, ataxin-7-92Q (1–266) or ataxin-7-92Q (1–266) K257R were co-transfected with vector control or HDAC3 in HEK293T cells. Western blot analysis of cellular lysates with ataxin-7 (PA1-749) antibody revealed the presence of bands corresponding to the wild type, expanded and ataxin-7 1–266 stop mutant. Bands of higher molecular weight were dominant when ataxin-7 was co-transfected with HDAC3 (indicated by asterisks). Note that these bands are diminished in the SUMOylation-deficient ataxin-7 mutant (K257R). β-actin was used as a loading control.