Mem eagle non essential amino acid solution

Colon adenocarcinoma cells DLD-1 (Horizon Discovery) were cultured in a 1:1 mixture of Roswell Park Memorial Institute-1640 medium (Sigma-Aldrich) and Ham's F-10 nutrient mix (Lonza), supplemented with 10% fetal calf serum (FCS; BioWest), and 1% penicillin/streptomycin (Sigma-Aldrich). Human embryonic kidney cells HEK-293T, osteosarcoma cells U2OS, hTERT-immortalized human fibroblasts VH10, SV40-transformed human fibroblasts C5RO, XP4PA (XPC-deficient) (19 (link)), CS1AN (CSB-deficient) (20 (link)), CS1AN complemented with YFP-CSB^del (21 (link)), and XRCC1-YFP expressing MRC-5 were cultured in a 1:1 mixture of Dulbecco's Modified Eagle medium (Lonza) and Ham's F-10, supplemented with 10% FCS, 1% penicillin/streptomycin, 2 mM UltraGlutamine (Lonza), and 0.1 mM MEM Eagle non-essential amino acid solution (Lonza). All cells were cultured at 37°C, in 20% O₂ and 5% CO₂, except for VH10 cells that were cultured at 3% O_2. To generate MRC-5 cells expressing XRCC1-YFP, XRCC1-YFP cDNA (a gift from Anna Campalans) (22 (link)) was cloned into pLenti-CMV-Puro-DEST (a gift from Eric Campeau & Paul Kaufman) (23 (link)). Following lentiviral transduction, MRC-5 cells stably expressing XRCC1-YFP were selected by puromycin and fluorescence-activated cell sorting.