Hd 105 007

All 2D cell lines used in this study were described previously: KB2P1.21, KP3.33 (Evers et al, 2008 (link)), DLD-1 BRCA2(−/−) (Horizon Discovery, #HD 105-007; RRID:CVCL_HD57), HEK293FT (RRID:CVCL_6911). For their culture, cell growth media were supplemented with 10% (v/v) fetal calf serum (FCS, Sigma) and 50 units/ml penicillin–streptomycin (Gibco). KB2P1.21 and KP3.33 cells were grown in Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12; Gibco) containing 5 μg/ml Insulin (Sigma, #I0516), 5 ng/ml cholera toxin (Sigma, #C8052) and 5 ng/ml murine epidermal growth factor(EGF, Sigma, #E4127). For the growth of the DLD-1 BRCA2(−/−) cells, the medium was additionally enriched with 2 mM l-glutamine and 25 mM sodium bicarbonate. HEK293FT cells were cultured in Iscove’s Modified Dulbecco’s Media (IMDM, Gibco) supplemented with 10% fetal calf serum (FCS, Sigma) and 50 units/ml penicillin–streptomycin (Gibco). Tissue culture was carried out under low oxygen conditions (37 °C, 5% CO₂, 3% O₂), except for the HEK293FT cells, which were cultured under standard conditions (37 °C, 5% CO₂). All cell lines used in this study are of female origin, except for DLD-1 BRCA2(−/−) cells (male). Testing for mycoplasma contamination was performed regularly, twice a year.

The human cell lines HEK293-T cells (gift from Dr. Mounira Amor-Gueret) were cultured in DMEM (Eurobio Abcys, Courtaboeuf, France) media containing 25 mM sodium bicarbonate and 2 mM l-glutamine supplemented with 10% FBS (EuroBio Abcys). The BRCA2-deficient colorectal adenocarcinoma cell line DLD1 BRCA2^{[−/−31 (link)} (HD 105-007) and the parental cell line DLD1 BRCA2^+/+ (HD-PAR-008) were purchased from Horizon Discovery (Cambridge, England). The cells were cultured in RPMI media (EuroBio Abcys) containing 25 mM sodium bicarbonate and 2 mM l-glutamine (EuroBio Abcys) Supplemented with 10% FBS (EuroBio Abcys). The DLD1 BRCA2^−/− cells were maintained in growth media containing 0.1 mg/ml hygromycin B (Thermo Fisher Scientific). The stable cell lines of DLD1^−/− BRCA2-deficient cells expressing BRCA2 WT or variants of interest generated in this study were cultured in growth media containing 0.1 mg/ml hygromycin B (Thermo Fisher Scientific) and 1 mg/ml G418 (Sigma-Aldrich).
All cells were cultured at 37 °C with 5% CO₂ in a humidified incubator and all cell lines used in this study have been regularly tested for mycoplasma contamination (MycoAlert, Lonza) and genotyped using GenePrint kit (Promega).