All mouse ESC lines used in this study were grown at 37 °C and 5% CO2 on gelatinised plates. Dulbecco’s Modified Eagle Medium (Gibco) was supplemented with 15% foetal bovine serum (Labtech), 2 mM L-glutamine (Life Technologies), 1x penicillin-streptomycin solution (Life Technologies), 1x non-essential amino acids (Life Technologies), 0.5 mM beta-mercaptoethanol (Life Technologies), and 10 ng/mL leukaemia inhibitory factor. Cells were passaged using trypsin-EDTA (0.25%, Gibco) with 2% chicken serum. Cells were regularly tested for the presence of mycoplasma.
To induce conditional knockout, PRC1CKO cells were treated with 800 nM 4-hydroxytamoxifen (OHT) (Sigma) for 72 h. To induce and maintain degradation and depletion of SUZ12, dTAG-SUZ12 were treated with 100 nM dTAG-13101 (link) for 96 h.
To induce conditional knockout, PRC1CKO cells were treated with 800 nM 4-hydroxytamoxifen (OHT) (Sigma) for 72 h. To induce and maintain degradation and depletion of SUZ12, dTAG-SUZ12 were treated with 100 nM dTAG-13101 (link) for 96 h.
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