Zeocin ant zn 1

hTERT-immortalized retinal pigmented epithelial cells (hTERT-RPE1, RRID:CVCL_4388) were cultured in standard growth conditions (5% CO2, 37°C) in complete media (1:1 Dulbecco Modified Eagle Medium / Nutrient Mixture F12 supplemented with 10% fetal bovine serum (FBS) (F9665, Merck), 1% penicillin-streptomycin (15140122, Life Technologies) and 0.123% sodium bicarbonate (S8761, Sigma-Aldrich)). hTERT-RPE1 Flp-In T-REx were grown under similar conditions, including 100 μg/ml Zeocin (ant-zn-1, InvivoGen) and 10 μg/ml Blasticidin (ant-bl-1, InvivoGen). hTERT-RPE1 Flp-In T-REx cells expressing RAD51 separation-of-function variants were generated by transfecting cells with pDEST_FRT_TO plasmids containing FLAG, FLAG-RAD51-WT, FLAG-RAD51-K133R, FLAG-RAD51-T131P or FLAG-RAD51-II3A. Selection was done in 500 μg/ml G418 (G8168, Sigma-Aldrich), and maintained in 200 μg/ml G418 and 10 μg/ml Blasticidin. To induce variant expression, cells were incubated with complete media containing 1 μg/ml doxycycline (D9891, Sigma Aldrich) for at least 24 hours. HCT116 cells were cultured in standard growing conditions (5% CO₂, 37°C) in McCoy’s 5A medium (26600080, Gibco) supplemented with 10% FBS and 1% penicillin-streptomycin. HeLa cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with high glucose (D6429, Sigma Aldrich) with 10% FBS and 1% penicillin-streptomycin.