Research Paper Volume 7, Issue 10 pp 839—853

Blocking the association of HDAC4 with MAP1S accelerates autophagy clearance of mutant Huntingtin

Figure 6. HDAC4 decreases levels of acetylated MAP1S. (A-D) HDAC4 reduces levels of acetylated MAP1S. HDAC4 was purified from 293T cells overexpressing HDAC4 by immunoprecipitation with antibody against HDAC4; purified HDAC4 (Active) was inactivated by boiling (Inactive). MAP1S purified from MAP1S overexpressing cells with MAP1S antibody was incubated with increasing amounts of active HDAC4 for 2 hrs (A, B) or same amount of HDAC4 for increasing amounts of time (C,D). Representative immunoblots (A, C) and quantification (B, D) are shown. (E, F) HDAC4 mutants with increased or reduced activity exhibit reduced or enhanced levels of acetylated MAP1S, respectively. HDAC4 mutant proteins purified in same way as the wildtype were incubated with purified MAP1S for 1 hr. Representative immunoblot results (E) and quantification (F) are shown.