Penicillin streptomycin glutamax

All cells were cultured at 37°C in a humidified 5 % CO₂ atmosphere. HEK293T cells (ATCC, female) used in transcriptional activation, NHEJ, and mCherry-disruption assays were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Life Technologies) supplemented with 10% fetal bovine serum (FBS, Sigma Aldrich) and 1× penicillin/streptomycin/glutamax (Life Technologies). U2OS.eGFP.PEST cells (human, female) stably integrated with an eGFP.PEST fusion gene were maintained in DMEM supplemented with 10% FBS, 1× penicillin/streptomycin/glutamax, and 400 μg/mL G418 (selection antibiotic). WM793 cells (human, male) were cultured in RPMI1640 (Life Technologies) media supplemented with 10% FBS and 1× penicillin/streptomycin/glutamax. Human islets (female) were maintained in CMRL 1066 (Life Technologies) containing 10% FBS, 1× penicillin/streptomycin/glut amax, and 1 mM sodium pyruvate and were cultivated at 37°C with 5% CO₂ in a humidified atmosphere. Human bone marrow-derived mesenchymal stem cells (hMSC, female) were cultured in DMEM supplemented with 10% FBS and 1× penicillin/streptomycin/glutamax. Cells were continuously maintained at <90% confluency. All cell lines were sourced commercially or were functionally validated. Cells were periodically tested for mycoplasma contamination using the MycoAlert PLUS Mycoplasma Detection Kit (Lonza).

All cells were cultured at 37 °C in a 5% CO₂ atmosphere. HEK293T cells (Life Technologies) used in transcriptional activation experiments were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (CellGro) supplemented with 10% FBS (CellGro) and 1% penicillin–streptomycin–glutamine (CellGro). HEK293T cells used in surveyor assays and nuclease specificity experiments and U2OS.eGFP-PEST cells^{26 (link)} stably integrated with an eGFP-PEST fusion gene were maintained in DMEM (Life Technologies) supplemented with 10% FBS, 1× penicillin–streptomycin–glutamax (Life Technologies) and 400 μg/mL of the selection antibiotic G418 (for the U2OS. eGFP-PEST cells). Cells were continuously maintained at <90% confluency. All cell lines were sourced commercially or were functionally validated. Cells were periodically tested for mycoplasma contamination using the MycoAlert PLUS Mycoplasma Detection Kit (Lonza).