Induced Tyrosine Phosphorylation of TH1 by EGF and Estrogen Stimulations.

<p>(A) Induced tyrosine phosphorylation of TH1 by EGF in HEK293T cells. HEK293T cells over-expressing GFP-TH1 were serum starved for 18 h, pretreated with 50 µM pervanadate for 15 min and incubated with or without 100 ng/ml EGF for 15 min in the presence of 10% fetal bovine serum. Cell lysates were immunoprecipitated with antibody against GFP. The tyrosine phosphorylation of TH1 was analyzed with pY (4G10). The same blot was striped and re-probed with anti-GFP antibody. (B) Induced tyrosine phosphorylation of TH1 by EGF in A549 cells. A549 cells were serum starved for 18 h, pretreated with 100 µM pervanadate for 15 min, stimulated with 100 ng/ml EGF for 5 min and 15 min in the presence of 10% fetal bovine serum. Cell lysates were immunoprecipitated with TH1 antibody. The tyrosine phosphorylation of TH1 was analyzed with pY (4G10). (C) Induced tyrosine phosphorylation of TH1 by Estrogen and EGF in MCF-7 cells. MCF-7 cells were serum and estrogen deprived, pretreated with 100 µM pervanadate for 15 min, and stimulated with10 nM 17β-estradiol (E2) for 2 min or 100 ng/ml EGF for 15 min in the presence of 10% fetal bovine serum. Tyrosine phosphorylation of TH1 was analyzed as in (B).