Ryu H, Lee J, Olofsson BA, Mwidau A, Dedeoglu A, Escudero M, Flemington E, Azizkhan-Clifford J, Ferrante RJ, Ratan RR, Deodoglu A. Histone deacetylase inhibitors prevent oxidative neuronal death independent of expanded polyglutamine repeats via an Sp1-dependent pathway. Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):4281-6. PubMed Abstract

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Immunoblot analysis was performed using Primary Abs against Sp1 (PEP2, Santa Cruz Biotechnology)diluted at 1:1,000 in 1% milk TBST and exposed to membranes overnight at 4°C. Immunoreactive proteins were detected according to the enhanced chemiluminescent protocol (Amersham Pharmacia). To monitor the Sp1 acetylation in vivo, neuronal lysates obtained from control and TSA-treated cells were prepared and run on SDS-page gels and proteins immunoreactive to acetyl lysine were detected by using an acetyl lysine-specific Ab (1:1,000 dilution; Upstate Biotechnology).