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. Author manuscript; available in PMC: 2014 Mar 14.
Published in final edited form as: Prostate. 2010 Sep 1;70(12):1274–1285. doi: 10.1002/pros.21163

Fig. 2.

Fig. 2

Generation of phospho-antibodies that specifically recognize pTyr267-AR. A: Serum and androgen-depleted LNCaP cells were treated with heregulin (10 ng/ml) for different time intervals and lysates were subjected to immunoblotting with pTyr267-AR antibodies (top panel) or with pTyr267-AR antibodies that were incubated with AR phospho267-peptide (second panel). B: Serumandandrogen-depleted LAPC4cells treated with EGF(10 ng/ml) for different time intervals. Lysates were immunoprecipitated with pTyr-antibodies, followed by immunoblotting with pTyr267-AR antibodies (top panel) or with pTyr267-AR antibodies that were incubated with AR phospho-peptide (second panel). C: HEK293 cells were transfected with the AR expression construct (2 μg) along with the caAck or kdAck expression construct (2 μg). Forty-eight hours after transfection lysates were immunoblotted with pTyr267-AR antibodies (top panel) or with pTyr267-AR antibodies that were incubated with AR phospho-peptide (second panel). D: Serum and androgen-depleted LNCaP and DU145 cells were treated with heregulin (10 ng/ml) for different time intervals. Lysates were immunoprecipitated with pTyr-antibodies, followed by immunoblotting with pTyr267-AR antibodies (top panel).