MCF-7 cells (from ATCC) were cultured in Minimal Essential Medium (Sigma) supplemented with 10% calf serum (HyClone), 100μg/mL penicillin-streptomycin (Invitrogen), and 25μg/mL gentamicin (Invitrogen). MDA-MB-157 cells were obtained from ATCC and cultured in Leibovitz’s L-15 medium (ATCC) supplemented with 10% fetal bovine serum (HyClone) and 100μg/mL penicillin-streptomycin (Invitrogen). Recombinant adenoviruses were constructed and prepared as described previously (7 (link)). Cells were infected with adenovirus expressing β-galactosidase (AdGal) or ERβ (AdERβ) as described (7 (link),10 (link)). The siRNA-mediated gene knockdowns (KD) were performed as described (23 (link)). ERα siRNA was purchased from Dharmacon. The siERα sequences are UCAUCGCAUUCCUUGCAAAdTdT and UUUGCAAGGAAUGCGAUGAdTdT. p53 siRNA was purchased from Santa Cruz (sc-29435).
Protocol detail
Breast Cancer Cell Culture and Adenoviral Infection
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MCF-7 cells
Minimal Essential Medium
penicillin-streptomycin
gentamicin
MDA-MB-157 cells
Leibovitz’s L-15 medium
p53 siRNA
Adenovirus
Adenoviruses
Cells
Cultured medium
Fetal bovine serum
Gene knockdowns
Gentamicin
Mcf 7 cells
Penicillin
Serum
Sirna
Streptomycin
β galactosidase
Corresponding Organization :
Other organizations : University of Illinois Urbana-Champaign
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Variable analysis
independent variables
- Adenovirus expressing β-galactosidase (AdGal) or ERβ (AdERβ)
- SiRNA-mediated gene knockdowns (KD) of ERα and p53
- Not explicitly mentioned
- MCF-7 and MDA-MB-157 cell lines
- Cell culture media (Minimal Essential Medium and Leibovitz's L-15 medium)
- Supplements (10% calf serum, 10% fetal bovine serum, 100μg/mL penicillin-streptomycin, 25μg/mL gentamicin)
- Positive control: Adenovirus expressing β-galactosidase (AdGal)
- Negative control: Not specified
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