Cell Culture Protocols for Various Cell Lines

Mouse embryonic stem cells were grown in Dulbecco’s Modified Eagle Medium (Thermo Fisher Scientific) supplemented with fetal bovine serum (FBS, 15% Biosera or 10% Sigma), 1× non-essential amino acids (Thermo Fisher Scientific), 2 mM l-glutamine (Thermo Fisher Scientific), 1× penicillin/streptomycin (Thermo Fisher Scientific), 0.5 mM beta-mercaptoethanol (Thermo Fisher Scientific), and 10 ng/ml leukaemia inhibitory factor (produced in-house). ESCs were grown on gelatinised plates at 37 °C and 5% CO₂. Cell lines expressing dTAG fusion proteins were treated with 100 nM dTAG-13 (produced by Behnam Nabet and Nathanael Gray^{87 (link)} or Carole Bataille and Angela Russell) to induce protein depletion.
Drosophila melanogaster SG4 cells were grown adhesively at 25 °C in Schneider’s Drosophila Medium (Thermo Fisher Scientific) supplemented with 1× penicillin/streptomycin and 10% heat-inactivated FBS (Biosera). Human HEK 293 T cells were grown at 37 °C and 5% CO₂ in Dulbecco’s Modified Eagle Medium, supplemented with 10% FBS (Biosera), 1× penicillin/streptomycin, 2 mM l-glutamine and 0.5 mM beta-mercaptoethanol. All cell lines were routinely tested to ensure they were mycoplasma free.