Hesr gfp

Cell lines, cultures, drug treatments, and plasmid transfections This study used breast cancer cell lines obtained from ATCC, including ERα+ (MCF-7 and T47D) and ERα-(MDA-MB-231). The MCF-7 cells were maintained in Dulbecco's Modi ed Eagle Medium (Cat No. 11995, Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (Cat No. 16000, Invitrogen, Carlsbad, CA) and 1% penicillin/streptomycin (Cat No. 15140, Invitrogen). The T47D and MDA-MB-231 cells were grown in Roswell Park Memorial Institute 1640 (Gibco, El Paso, TX) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. All cells were cultured at 37°C in the presence of 5% CO 2 . The cells were starved for 24 hours and treated with ICI (fulvestrant, Sigma-Aldrich, St. Louis, MO, 1 to 10 µM), IFN-γ (R&D Systems, Minneapolis, MN, 100 units/ml), or β-estradiol (Sigma-Aldrich, 1 nM) in 2 ml of medium for an appropriate time. The cells were then used in the protein expression assays.
For the ESR1 plasmid transfection, ERα-cells were plated and cultured in a 6-well plate at 90% con uency and transfected with 2.5 µg of hESR-GFP (Cat No. #28230, Addgene, Cambridge, MA) using 3.75 µl of Lipofectamine ® 3000 reagent (Life Technologies) and 5 µl of P3000™ reagent (Life Technologies) per well according to the manufacturer's protocol.

Cell lines, cultures, drug treatments, and plasmid transfections This study used breast cancer cell lines obtained from ATCC, including ERα+ (MCF-7 and T47D) and ERα-(MDA-MB-231). The MCF-7 cells were maintained in Dulbecco's Modi ed Eagle Medium (Cat No. 11995, Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (Cat No. 16000, Invitrogen, Carlsbad, CA) and 1% penicillin/streptomycin (Cat No. 15140, Invitrogen). The T47D and MDA-MB-231 cells were grown in Roswell Park Memorial Institute 1640 (Gibco, El Paso, TX) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. All cells were cultured at 37°C in the presence of 5% CO 2 . The cells were starved for 24 hours and treated with ICI (fulvestrant, Sigma-Aldrich, St. Louis, MO, 1 to 10 µM), IFN-γ (R&D Systems, Minneapolis, MN, 100 units/ml), or β-estradiol (Sigma-Aldrich, 1 nM) in 2 ml of medium for an appropriate time. The cells were then used in the protein expression assays.
For the ESR1 plasmid transfection, ERα-cells were plated and cultured in a 6-well plate at 90% con uency and transfected with 2.5 µg of hESR-GFP (Cat No. #28230, Addgene, Cambridge, MA) using 3.75 µl of Lipofectamine ® 3000 reagent (Life Technologies) and 5 µl of P3000™ reagent (Life Technologies) per well according to the manufacturer's protocol.