FIG. 8.
(A) TSA enhances CIITA recruitment to the MHC-II promoter. 293T cells were transfected with Fg-CIITA and treated with 300 nM TSA. Chromatin immunoprecipitation was performed using anti-Fg M5. MHC-II promoter DNA was detected by quantitative real-time PCR. Data are presented as increases compared to results with untreated cells. Real-time PCR values were determined by subtracting values obtained from bead-only immunoprecipitations and normalizing to the total amount of MHC-II promoter DNA added to the immunoprecipitation reaction (input). Data shown are representative of three independent experiments. (B) TSA does not promote association of CIITA with the β-actin promoter. Chromatin was prepared from transiently transfected 293T cells as was done for panel A. PCR was performed to detect β-actin promoter DNA sequences. Input represents 1% of the total chromatin introduced into each immunoprecipitation reaction. (C) TSA does not affect CIITA protein levels. 293T cells were transfected with Fg-CIITA and treated with TSA as for panel A. CIITA was detected by Western analysis anti-Fg antibody. As a loading control we also immunoblotted with antibodies against actin (bottom panel).