Figure S5_S6 from Inhibin Is a Novel Paracrine Factor for Tumor Angiogenesis and Metastasis

Figure S5: Representative western blot of 6 hr serum starved HMEC-1 cells treated with CM from either shControl or shINHA SKOV3 cells. Anti-INHA (polyclonal; 10 µg/ml; #sc22048, Santa Cruz) antibody was used as indicated. Samples were immunoblotted for phospho-Smad1/5, total Smad1/5 and GAPDH. Quantitation from two independent trials is presented (right graph). Figure S6: (i) Western blotting of lysates from HMEC-1 (above) or MEECs (below) incubated with recombinant 20pM Inhibin A for indicated times or increasing doses of recombinant Inhibin A as indicated for 30 min followed by immunoblotting. (ii) Smad1/5 phosphorylation was analyzed in response to 20 pM Inhibin A over time, or increasing doses of Inhibin A at a fixed time point of 30 min in SKOV3 epithelial cancer cells. Lysates were immunoblotted for phospho-Smad1/5, Smad1/5 and GAPDH. HMEC-1 cells with 100pM Inhibin A treatment were tested side-by-side for comparison.