Human WT- ER+ BC cells MCF7, ZR75, and T47D, human mammary epithelial cells (HMEC), murine mammary epithelial cells (HC11), primary human endometrial epithelial cells (HEEC), primary human endometrial stromal cells (HESC), and primary human nontumorigenic immortalized ovarian surface epithelial cells (IOSE-80) were obtained from the American Type Culture Collection (ATCC) and were maintained using ATCC recommended media. MCF7 and ZR75 model cells stably overexpressing PELP1 cDNA or PELP1-shRNA have been described previously (18 (link)). MCF7-TamR (tamoxifen resistant) and MCF7-LTLT (letrozole resistant) cells were cultured in tamoxifen or letrozole (1 μM) containing media (19 ). MCF7(Y537S) and MCF7(D538G) MT- ER+ BC cell lines were described previously (20 (link)). ZR75(Y537S) and ZR75(D538G) MT- ER+ BC cell lines were generated in our lab (19 ). All model cells utilized were free of mycoplasma contamination. Additionally, STR DNA profiling was used to confirm the identity of cells. The GAPDH (8884) antibody was obtained from Cell Signaling Technology (Beverly, MA). The β-Actin (A-2066) and Vinculin antibodies (V9264) were purchased from Millipore Sigma (Burlington, MA). The Ki67 antibody (ab1667) was purchased from Abcam (Cambridge, MA). The PELP1 antibody (A300-180A) was purchased form Bethyl Laboratories Inc. (Montgomery, TX).
Protocol detail
Characterization of Breast Cancer Cell Lines
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Actin
Antibodies
Antibody
Cdna
Cell lines
Culture media
Endometrial
Epithelial cells
Gapdh
Hc11
Human
Letrozole
Mammary
Mcf7
Murine
Mycoplasma
Ovarian
Pelp1
Shrna
Stromal cells
Tamoxifen
Vinculin
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Variable analysis
independent variables
- PELP1 cDNA overexpression
- PELP1-shRNA knockdown
- Tamoxifen treatment
- Letrozole treatment
- Estrogen receptor (ER) mutations (Y537S, D538G)
- Cell line characteristics and behavior
- Cell culture media and conditions
- Cell type/origin (WT-ER+ BC cells, HMEC, HC11, HEEC, HESC, IOSE-80)
- Mycoplasma contamination status
- Cell line identity (STR DNA profiling)
- Positive control: WT-ER+ BC cell lines (MCF7, ZR75, T47D)
- Negative control: Not explicitly mentioned
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