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. 2010 May 3;189(3):407–415. doi: 10.1083/jcb.201001148

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© 2010 Zheng et al.

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Figure 3. — pS187-H1.4 is preferentially associated with transcriptionally active rDNA. (A) The levels of pS187-H1.4 and total H1.4 (α-H1.4) at the 45S pre-rRNA promoter in untreated and 50 ng/ml ActD-treated HeLa cells (3 h). The data are expressed as fold change relative to a parallel ChIP without primary antibody from untreated cells. (B) The levels of the 45S pre-rRNA transcript in untreated and ActD-treated cells were assayed by RT-PCR and normalized to β-actin expression. The data in A and B represent the mean of two independent experiments. Error bars indicate the standard error of the mean for triplicate samples. (C) The global levels of pS187-H1.4 and total H1.4 in ActD-treated cells were assayed by immunoblotting and quantified relative to the control samples after normalization to α-tubulin levels. MW, molecular weight. (D) Confocal images of control and ActD-treated HeLa cells costained with antibodies to pS187-H1.4 and fibrillarin, UBF, or the large subunit of RNA polymerase I (RPA 194). DNA was visualized using TO-PRO-3. Bars, 5 µm.