BMP-2 and Wnt3a promote proliferation, migration, and survival of hPAECs while increasing β-C transcriptional activity and target genes. (A) Cell count, caspase 3/7 apoptosis, and microcarrier bead migration assays of hPAECs in the presence or absence of 10 ng/ml BMP-2 or 100 ng/ml Wnt3a. Serum-free conditions were used to induce apoptosis, whereas 50 ng/ml of recombinant human VEGF was used as a positive control for proliferation and motility. (B) Representative Western immunoblots (top) and densitometry values (bottom) for β-C normalized to α-tubulin in response to BMP-2 and Wnt3a stimulation as in A. (C) TOPflash luciferase assays. hPAECs were transfected with TOPflash or FOPflash (negative control) luciferase reporter plasmids or Renilla (control for transfection efficiency). 6 h after stimulation with BMP-2 and Wnt3a, lysates were analyzed for luciferase activity relative to Renilla. CON, control. (D) Representative immunoblots for c-myc, VEGF, cyclin D1, and survivin in lysates of hPAEC stimulated with Wnt3a or BMP-2 as described in A. Values were normalized for α-tubulin. Error bars represent mean ± SEM from four different experiments performed in triplicate. *, P < 0.01; **, P < 0.001; ***, P < 0.0001 (vs. unstimulated control).