Ets1 cooperates with ERα and nuclear receptor coactivators to stimulate an estrogen responsive promoter.

<p>A. HeLa cells were transiently transfected with 75 ng ERE2-TK-LUC, 0.2 ng pcDNA3.1-hERα and 50 ng pSG5-hEts1 or control vector to hold DNA concentration constant in Optimem serum free media. After 24 hours cells were treated with 10 nM estradiol where indicated. Samples were harvested and analyzed after 24 hours of treatment. Bars represent mean ± SEM of triplicate samples. * Different from ERα alone (bar 5) p<0.05. B. HeLa cells were transiently transfected and treated as in A with the addition of 25 ng of pCR3.1 NCOA expression constructs or control vector. Bars represent mean ± SEM of triplicate samples. * Different from ERα with each respective NCOA alone (bars 5, 13, 21) p<0.05. C. Western blot analysis of ERα expression in transfected cells. Transfected cells were harvested and 100 µg of total protein was analyzed for ERα expression by Western blot. The blot was stripped and probed with an antibody against tubulin to control for loading.