FIGURE 7.

HMGN2 does not affect post-translational histone modifications. A, HMGN2 does not affect histone acetylation at H3K9 or H3K14. T47D cells expressing shCTL or shHMGN2 were treated with or without PRL for 45 min and were analyzed by ChIP-qPCR at the CISH promoter. Nuclear lysates were precipitated with antibodies against acetylated H3K9 (H3K9Ac), H3K14 (H3K14Ac), or total H3. CISH primers are described in the legends to Figs. 2 and 5A. The percentage of input DNA recovered with the acetyl-specific antibodies was normalized to the percentage of input recovered for total H3 in a parallel sample. Results are presented as the mean ± S.E. (error bars), n ≥ 3 independent experiments. Statistical significance was determined by two-way repeated measures ANOVA. B, the CISH promoter is not enriched for histone trimethylation at H3K9 or H3K27. ChIP-qPCR analysis was carried out as in A using antibodies against H3K9me3 or H3K27me3. Positive (+) controls for enrichment are ZNF554 for H3K9me3 and α-Satellite for H3K27me3 (EMD Millipore). The amount of DNA recovered was calculated relative to the input control and is graphed as a percentage of input. Results are presented as the mean ± S.E. of three independent experiments. Statistical significance was determined by two-way ANOVA. n.s., p > 0.05; *, p ≤ 0.05; **, p ≤ 0.01.