Journal: Breast Cancer Research
Article Title: Phosphorylation of estrogen receptor α serine 167 is predictive of response to endocrine therapy and increases postrelapse survival in metastatic breast cancer
doi: 10.1186/bcr1285
Figure Lengend Snippet: Immunoblot analysis of phosphorylated estrogen receptor (ER)-α Ser118 and ER-α Ser167. (a) Transfected COS-7 cells were grown in serum- and estrogen-deprived conditions and treated with vehicle (medium) (lane 1), 17β-estradiol (E 2 ) (lane 2), epidermal growth factor (EGF) (lane 3), or E 2 and EGF (lane 4) for 30 min. Equal amounts of total protein from whole cell lysates were blotted for either anti-ER-α-phosphoserine (α-pS118 and α-pS167) and anti-ER-α (α-ER-α) antibodies. (b) T47D cells were grown in serum- and estrogen-deprived conditions and treated with vehicle (medium) (lane 1), 17β-estradiol (E 2 ) for 10 min (lane 2) and 30 min (lane 3), EGF for 10 min (lane 4) and 30 min (lane 5), or E 2 and EGF for 10 min (lane 6) and 30 min (lane 7). Equal amounts of total protein from whole cell lysates were blotted for either anti-ER-α-phosphoserine (α-pS118 and α-pS167) and anti-ER-α (α-ER-α) antibodies.
Article Snippet: Primary antibodies included monoclonal mouse antihuman ER-α antibody (1D5, DAKO, Glostrup, Denmark) at 1:100 dilution for ER-α; polyclonal rabbit antiphospho-ER-α (Ser118) antibody (Cell Signaling) at 1:25 dilution for phosphorylated ER-α Ser118; polyclonal rabbit antiphospho-ER-α (Ser167) antibody (Cell Signaling) at 1:50 dilution for phosphorylated ER-α Ser167; polyclonal rabbit anti-ER-β1 antibody at 1:10000 dilution for ER-β1; polyclonal rabbit anti-ER-βcx/β2 antibody at 1:2000 dilution for ER-βcx/β2; monoclonal mouse antihuman PR antibody (636, DAKO) at 1:100 dilution for PR; monoclonal mouse antihuman PR antibody (Ab-7, Neo Markers, Fremont, CA) at 1:100 dilution for PRA; and monoclonal mouse antihuman PR antibody (Ab-2, Neo Markers) at 1:100 dilution for PRB.
Techniques: Western Blot, Transfection