Mda 231

MDA-MB-157 (MDA-157), MDA-MB-231(MDA-231) and MDA-MB-468 (MDA-468), MCF10A, and HCC1806 cells were purchased from the American Type Culture Collection (ATCC #’s HTB-24, HTB-26, HTB-132, CRL-10317, and CRL-2335, respectively) within the last 12 months and passaged < 15 times for all experiments. Cells were tested biweekly during experiments for mycoplasma contamination using the Lonza MycoAlert (Lonza #LT07-318). HCC1806 cells were grown in RPMI 1640 Medium (Life Technologies #11875093) and supplemented with 10% Fetal Bovine Serum (FBS, Atlantic Biologicals Premium Select). MDA-157, MDA-231, MDA-468, and MCF10A cells were grown in DMEM High Glucose + GlutaMAX (Life Technologies #10566016) and supplemented with 1% sodium pyruvate (Life Technologies #11360070) and 10% FBS. Cells were maintained in a humidified 37°C incubator with 5% carbon dioxide.

The TNBC cell lines HCC1806, HCC1937, MDA-MB-157 (MDA-157), MDA-MB-231 (MDA-231), and MDA-MB-468 (MDA-468) were purchased from the American Type Culture Collection (Manassas, VA; CRL-2335, CRL-2336, HTB-24, HTB-26, and HTB-132). MX-1 was purchased from the NCI repository. All cell lines were purchased within the previous 24 months and passaged < 15 times for all experiments [Table 1]. Cells were tested biweekly for mycoplasma contamination (MycoAlert, Lonza, Basel, Switzerland). MX-1, MDA-157, MDA-231, and MDA-468 were grown in Dulbecco Modified Eagle Medium (DMEM High Glucose with GlutaMAX, Life Technologies, Carlsbad, CA) and supplemented with 1% sodium pyruvate (Life Technologies) and 10% fetal bovine serum (FBS) (Premium Select, Atlantic Biologicals, Miami, FL). HCC1937 and HCC1806 were grown in RPMI supplemented with 10% FBS. Cells were maintained in a humidified 37 °C incubator with 5% carbon dioxide.

MDA-MB-231 (MDA231), MCF7 and HEK293T cells were obtained from ATCC (The Global Bioresource Center). MCF7 cells were cultured in Roswell Park Memorial Institute 1640 (RPMI1640, Life Technologies, USA) medium supplemented with 10% fetal bovine serum (FBS). MDA231 and HEK293T cells were routinely cultured in Dulbecco’s modified Eagle’s medium (DMEM, Life Technologies, USA) containing 10% FBS. Transfection was performed with Lipofectamine 3000 according to the manufacturer’s protocols (Invitrogen, USA), as described previously [12 (link)]. The BRD7-overexpressing and BRD7-shRNA cells were generated by lentiviral infection. BRD7-overexpressing lentivirus was purchased from GenePharma (Suzhou, China), YB1-overexpressing lentivirus was obtained using a YB1 expression plasmid purchased from Sino biological (Beijing, China) and packaged in HEK293T cells, and BRD7 shRNA lentivirus was obtained using the expression vector pLVTH/shBRD7. The BRD7 siRNA sequence was 5′-GUGCCAAGAUUAUCCGUAUdTdT-3′. A total of 10 μg of the corresponding expression vector and 7.5 μg of the packaging vectors (pMD2G and pSPAX2) were co-transfected into HEK293T cells for 48 h. The virus-containing supernatant was collected, centrifuged at 2000 rpm for 10 min and filtered through a 0.22 μm membrane. Tumor cells were infected with the supernatant for 48 h and screened for 72 h with 2 μg/ml puromycin in DMEM.

MDA-MB-231(MDA-231), MDA-MB-468 (MDA-468), and HEK293T were purchased from the American Type Culture Collection (ATCC HTB-26, HTB-132, and CRL-3216, respectively; Manassas, VA, USA) within the last 24 months and passaged < 15 times for all experiments. Cells were tested biweekly during experiments for mycoplasma contamination using the Lonza MycoAlert^® (Lonza #LT07-318). MDA-231 and MDA-468 cells were grown in DMEM High Glucose + GlutaMAX™ (Life Technologies, Carlsbad, CA, USA, #10566016) and supplemented with 1% sodium pyruvate (Life Technologies, #11360070) and 10% FBS (Premium Select, R&D systems, Minneapolis, MN, USA). HEK293T cells were grown in DMEM High Glucose + L-Glutamine (HyClone, Logan, UT, USA, # SH30022.01) and supplemented with 1% sodium pyruvate (Life Technologies #11360070) and 10% FBS. Cells were maintained in a humidified 37 °C incubator with 5% carbon dioxide.

MDA-MB-231 (ATCC cat#: HTB-26) and MCF10A (ATCC cat#: CRL-10317) were purchased from American Type Culture Collection (ATCC, Manassas, VA). MCF7 cells (ATCC cat#: HTB-22) were obtained from Mary Alpaugh’s lab (Rowan University, Camden NJ). The MDA-231 and MCF7 cells were grown in Dulbecco’s Modified Eagle Medium + GlutaMAX (Gibco cat#:10566024) with 10% Fetal Bovine Serum (Gibco, cat#: 16140089) and 1% Penicillin/Streptomycin. MCF10A cells were grown in Ham’s F-12 (Modified) + L-glutamine media (Corning, Cat#: 10-080-CV) with 5% horse serum (Gibco/Life Technologies, cat#: 16050-130), 0.5 mg/ml hydrocortisone (ThermoFisher Scientific, Waltham, MA, cat#: AC35245-0010), 100 ng/ml cholera toxin (MilliporeSigma, Burlington, MA, cat#: 227036), 10 μg/ml Insulin (Sigma-Aldrich, St. Louis, MO, cat#: 10516), and 1% Penicillin/Streptomycin. All cells were grown at 37°C and 5% CO₂. Mycoplasma testing is done bimonthly using the MycoStrip 100 kit (InvivoGen, cat#: rep-mysnc-100).

Human mammary epithelial (MCF10A), immortalized mouse embryonic fibroblast (NIH3T3) and breast cancer cell lines (MCF-7, T47D, MDA-231 SKBR3) were procured from ATCC MCF10A cells were cultured and maintained in a 1:1 mixture of Dulbecco's Modified Eagle Medium and Nutrient Mixture F-12 medium (DMEM/F12) (Gibco, Carlsbad, CA) supplemented with 5% horse serum (HS), 0.5 μg/ ml hydrocortisone (Sigma-Aldrich, St. Louis, MO, USA), 20ng/ ml epidermal growth factor EGF (Peprotech, Rocky Hill, NJ, USA), and 10 μg/ ml insulin (Sigma-Aldrich, St. Louis, MO, USA) and 100 ng/ ml cholera toxin (Sigma-Aldrich). NIH3T3, MCF-7, T47D, MDA-231 and SKBR3 cell lines were cultured and maintained in DMEM (Gibco, Carlsbad, CA) supplemented with fetal bovine serum (Gibco) to a final concentration of 10%, 1% Pen/Strep (Gibco), and 1% L-Glutamine (Gibco).The entire cell lines were cultured in CO2 (5%) incubator at 37 °C with 95% humidity.