Figure 1

Activation of Src and STAT3 in OSA cell lines and tissues. A) OSA cells and normal canine osteoblasts were serum starved then left untreated or stimulated with rhHGF (50 ng/ml). Protein lysates were generated and separated by SDS PAGE and Western blotting for pSTAT3 (Y705), pSrc (Y416), total Src, and total STAT3 was performed. B) OSA cell lines were serum starved for 2 hours while in suspension culture (S), serum starved for 2 hours while remaining adherent to tissue culture flasks (A), or left in 1% serum while adherent in flasks for 2 hours prior to collection (C). Protein lysates were generated and separated by SDS-PAGE and Western blotting for pSTAT3 (Y705), pSrc (Y416), total Src, and total STAT3 was performed. C) Fresh frozen canine OSA tumor tissues and control normal muscle tissue were processed for protein lysates. Protein was separated by SDS-PAGE and Western blotting for pSTAT3 (Y705), STAT3, VEGF, and β-actin was performed. D) RNA was collected from canine OSA cell lines and RT-PCR was performed for SFK members Src, Fyn, Yes, and Lyn as well as GAPDH as a control.