Figure 8. SMARCA4 is required for SMAD2 binding, nucleosome eviction and histone acetylation at a subset of Activin target genes.

(A) P19 cells were transfected with either non-targeting (NT) or Smarca4 siRNAs. Cells were then signal inhibited (SB-431542) or stimulated with Activin for 1 hr after SB-431542 washout. They were assayed for transcription by qPCR. The data shown are means ± SEM from four independent experiments. (B–D). Samples were prepared as for A and assayed for SMAD2 ChIP-PCR (B), H3K27Ac ChIP-PCR (C), or FAIRE-PCR (D) on the selected SBSs indicated. Plotted in B are the means and SEM of four independent experiments. Neg ctrl, negative control. The data in C and D are from a representative experiment of three (means ± SD). See Figure 8—figure supplement 1 for the averages of the three experiments and the statistical analyses. In A and B, ^**** corresponds to a p value of < 0.0001. ^** corresponds to a p value of < 0.01; n.s., not significant.

DOI: http://dx.doi.org/10.7554/eLife.22474.027

Figure 8—figure supplement 1. The role of SMARCA4 in SMAD2-mediated chromatin remodeling.

(A and B) P19 cells were transfected with either non-targeting (NT) or Smarca4 siRNAs, and were then signal inhibited (SB-431542), or stimulated with Activin for 1 hr after SB-431542 washout. The nucleosomes flanking or at selected SBSs were assayed for H3K27Ac enrichment by ChIP-PCR (A) or assayed for FAIRE-PCR (B). Plotted are the means and SEM of three independent experiments performed in duplicate. n.s., not significant. **** corresponds to a p value <0.0001, *** corresponds to a p value of < 0.001, ** corresponds to a p value of < 0.01.

DOI: http://dx.doi.org/10.7554/eLife.22474.028