Mccoy s 5a

Three cell lines representing different molecular subtypes of breast cancer were enrolled in the study, i.e., (i) MCF7 (ER/PR+, HER2low, TP53WT), (ii) MDA MB-231—basal-like subtype, also called triple-negative breast cancer (TNBC; ER/PR-, HER2-, TP53mut), and (iii) SK-BR-3 (ER/PR-, HER2+, TP53mut). MCF7 cell line, in comparison to the MDA-MB-231 cell line, is a poorly aggressive and non-invasive cell line. Overall, it is being considered to have low metastatic potential. SK-BR-3 cells are the least invasive cells out of the three studied, according to previous findings [22 ]. The MCF7 (HTB-22) and MDA-MB-231 (HTB-26) cells were maintained in RPMI-1640 (Biowest, Nuaillé, France), while SK-BR-3 (HTB-30) cells were cultured in McCoy’s 5A (Biowest, Nuaillé, France) media supplemented with 10% fetal bovine serum (FBS) (Biowest, Nuaillé, France) at 37 °C in an atmosphere of 5% CO₂ and saturated humidity. All cell lines were obtained from the American Type Culture Collection (ATCC).

Immortalized Bax−/− Bak−/− (DKO) MEFs and their wild type (MEFs) counterparts were obtained from Dr. Korsmeyer’s laboratory. Fip200−/− MEFs and their counterparts (Fip200+/+ MEFs) were gently supplied by Dr. Molinari and originated at Dr. Guan’s laboratory. SH-SY5Y, HeLa, HEK 293 cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA). HCT116 human adenocarcinoma cell line was kindly provided by Dr Vogelstein (The Howard Hughes Medical Institute, Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins School of Medicine, Baltimore, MD, USA).
All cell lines were maintained in DMEM (Gibco, Paisley, Scotland, UK) supplemented with 10% FCS (Gibco, Paisley, Scotland, UK) with the exception of HCT116, which were maintained in McCoy’s 5A (Biowest, Riverside, MO, USA) supplemented with 10% FCS. 5 μg/ml Plasmocin™ (InvivoGen, San Diego, CA, USA) was used as the media antibiotic. General culturing conditions were 37°C and a water-saturated, 5% CO₂ atmosphere. Culture dishes and other plastic disposable tools were supplied by VWR (Radnor, PA, USA) and Becton Dickinson (Franklin Lakes, NJ, USA).

BxPC-3 and Capan-1 cell lines were purchased from the ATCC (Manassas, VA, USA). MIA PaCa-2, MCF7, SK-BR-3 and MDA-MB-231 cell lines were in possession of 4Cell Therapies S.A. and were authenticated by Eurofins Genomics (Ebersberg, Germany). Capan-1 cells were cultured in IMDM, 20% FBS and 1× pen/strep (Biowest) on collagen I (Merck, Darmstadt, Germany)-coated plates. The BxPC-3 cell line was cultured with RPMI 1640 with 10% FBS and 1 × pen/strep (Biowest). The SK-BR-3 cell line was cultured in McCoy’s 5a supplemented with 20% FBS and 1 × pen/strep (Biowest). All other cell lines were cultured in DMEM high glucose supplemented with 10% FBS and 1 × pen/strep (Biowest).