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. 2004 Nov 9;101(47):16537–16542. doi: 10.1073/pnas.0404429101

Fig. 4.

Fig. 4.

Recruitment of α-IκBα correlates with histone deacetylation and hes1 repression. (a) Chromatin prepared from TNF-α-treated 3T3 cells was immunoprecipitated with α-IκBα (Upper) and α-AcH3-K14 (Lower) antibodies. Coprecipitated DNA was analyzed by PCR with specific primers for hes1 or histone H4. Results are representative of two independent experiments. (b) TNF-α treatment induces a temporary activation of the endogenous hes1 transcription. Northern blot from 293T cells treated with TNF-α at different time points showing hes1 mRNA levels. 28s rRNA is shown as a loading control. (c) Chromatin from control or 60 min TNF-α-treated 3T3 cells was first precipitated with α-IκBα (Lower). Chromatin was then eluted and a second precipitation with α-HDAC1 or -5 antibodies was performed (Upper). The presence of hes1 promoter in the precipitates was determined by PCR analysis. (d) Northern blot showing relative hes1 transcriptional activation in IκBα+/+ (Left) and IκBα–/– (Right) MEF treated with TNF-α at the indicated time points. 28s ribosomal RNA is shown as a loading control. RNA levels were quantified, and the ratio between hes1 and 28s is represented (Lower). Induction of IκBα transcription in the IκBα+/+ is shown as a control of TNF-α activation (Lower).