Figure 5: SMAD6 mediates Notch-dependent suppression of BMP signalling.
From: Notch regulates BMP responsiveness and lateral branching in vessel networks via SMAD6
(a) ChIP assay on micrococcal nuclease-digested HUVEC DNA immunoprecipitated with indicated antibodies and amplified unique primers targeting a single putative RBPJ consensus sequence in the SMAD6 promoter. Left, size marker (bp). (b–i) HUVEC transfected with non-targeting (b,c,f,g) or SMAD6 siRNA (d,e,h,i), plated onto IgG (b,d,f,h) or Dll4-Fc (c,e,g,i), then treated with control (b–e) or 50 ng ml−1 BMP6 (f–i) for 90 min before staining for nuclear pSMAD1/5. Scale bar, 10 μm. (j) Quantification of nuclear pSMAD1/5 fluorescence intensity, representative of three independent experiments. Data points, individual nuclei (n indicated on graph); bars, mean±95% CI. *P≤0.05, **P≤0.01 and ****P≤0.0001 by Kruskal–Wallis with Dunn’s post-hoc test.
