Figure 1.

HDAC activity is required for PU.1 gene expression. A) MEL cells were treated with HDACi [100 nM TSA or 10 nM depsipeptide (Depsi)] for 8 h. PU.1 mRNA level was measured by using real-time PCR. B) K562 cells were treated with an increasing amount of TSA with or without 20 μm cycloheximide (CHX). PU.1 mRNA level was measured by using real-time PCR. C) K562 cells were treated with 10 nM Depsi for different time points. PU.1 mRNA level was measured by using real-time PCR. D) MEL cells were induced with 1.5% DMSO at different time points, and PU.1 mRNA level was measured by using real-time PCR. E) Recruitment of HDAC1 and acetyl-HDAC1 on mouse PU.1 promoter. ChIP assays with HDAC1 and acetylated HDAC1 Ab was performed in MEL cells induced with DMSO for 0 and 3 d. The resulting precipitated DNA was analyzed by using real-time PCR. F) Human CD34⁺ cells were induced for differentiation with Epo for 5 d, and CD36⁺ cells were collected via flow cytometry after differentiation. PU.1 mRNA level was measured by using real-time PCR. G) Human CD34⁺ cells were treated with TSA for 8 h. PU.1 mRNA level was measured by using real-time PCR. H) HDAC1 recruitment on human PU.1 promoter. ChIP assays with HDAC1 Ab were performed in CD34⁺ or CD36⁺ cells. The resulting precipitated DNA was analyzed by using real-time NT, no treatment; TSS, transcription start site; UTR, untranslated region. All data are represented as means ± sem.