Figure 5. PDGF blocks PTEN and SRF binding to CArG elements in SM genes.

SMCs were serum-restricted for 48 h followed by stimulation with vehicle control or 20 ng ml⁻¹ platelet-derived growth factor-BB (PDGF-BB) for an additional 48 h. (a) Representative western blot for total PTEN, phospho-Akt, SRF and αSMA; N>3 independent experiments. (b–d) ChIP assays for PTEN (b) and SRF (c,d) binding to CArG elements in the Acta2 (b,c) and Fos (d) promoters were performed as described above. Data represent average fold changes±s.e.m. N=3 independent experiments; unless otherwise noted, *P<0.01 versus vehicle control; ‘**' denotes statistical analysis not conducted due to complete loss of PTEN interactions in PDGF-stimulated SMCs (consistent undetectable values). Molecular weight markers were cropped out for final SRF blots; please see Supplementary Fig. 8.