Fetal bovine serum (fbs)

The DT40 cell line is derived from chicken B lymphoma [22 (link)] and was cultured at 39.5°C with 5% CO₂ in RMPI-1640 medium (Nacalai Tesque, Kyoto, Japan) with glutamine (11875, Invitrogen, US) supplemented with 1% chicken serum (GIBCO-BRL, Grand Island, NY, USA), 10% heated-inactivated fetal bovine serum (FBS) (100–106, Gemini Bio-Products, West Sacramento, CA), 50 μM mercaptoethanol (Invitrogen), 50 U/ml penicillin and 50 μg/ml streptomycin (Nacalai Tesque). Doxycycline was added at a final concentration of 100 ng/ml to inactivate the expression of the tetracyclin repressible promoter.
Human cell lines (obtained from American Type Culture Collection, ATCC) were maintained at 37°C with 5% CO₂ in DMEM media (Nacalai Tesque) supplemented with 10% FBS (Gemini Bio-Products) for U2OS and MCF7 cell lines or in RPMI-1640 (Nacalai Tesque) with 10% FBS (Gemini Bio-Products) for HCC1937 cell lines.

CAL148 and MDA-MB-453 were grown in DMEM (Thermo Fisher Scientific, 11965–092) with 1 μg/mL EGF (Thermo Fisher Scientific, PHG0313) and 10% FBS (Gemini Bio Products, 100–106). HCC1806 (ATCC, CRL-2335) and HCC70 (ATCC, CRL-2315) cells were grown in RPMI + GlutaMAX (Thermo Fisher Scientific, 61870–036) with 10% FBS (Gemini Bio Products, 100–106). MCF10 (ATCC, CRL-10317) and HMEC cells were grown as previously described [83 (link)]. HaCaT (Cell Line Services, 300493), HaCaT C2 (single cell clone derived from parental HaCaT), HaCaT p63-/- (lack p63α expression as a result of CRISPR-Cas9 genomic editing of HaCaT C2 cells), 293FT (Thermo Fisher Scientific, R70007), and MDA-MB-231 (ATCC, HTB-26) cells were grown in DMEM (Thermo Fisher Scientific, 11965–092) with 10% FBS (Gemini Bio Products, 100–106). HDFn (ATCC, PCS-201-010) cells were grown in Medium 106 (Thermo Fisher Scientific, M-106-500) with 2% (v/v) Low Serum Growth Supplement (Thermo Fisher Scientific, S-003-10). All cell lines were grown in 100 U/mL Penicillin:Streptomycin (Gemini Bio Products, 400–109) and tested negative for mycoplasma (Lonza, LT07-418). HDFn cells were passaged using Trypsin-EDTA for Primary Cells (ATCC, PCS-999-003) and Trypsin Neutralizing Solution (ATCC, PCS-999-004). All other cells were passaged using 0.25% Trypsin-EDTA (Thermo Fisher Scientific, 25200–056).

HCT116, H1299, SK-HEP1, and U2OS cells were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum (Gemini Bioproducts, cat# 900-108). HT-1080 cells were maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum (Gemini Bioproducts, cat# 900-108), and 1% non-essential amino acids (Sigma-Aldrich, cat# M7145). RKO cells were grown in McCoy’s 5A modified medium (Gibco, cat# 16600-082) supplemented with 10% heat-inactivated fetal bovine serum (Gemini Bioproducts, cat# 900-108). The HCT116 and RKO isogenic cell lines that were created by the deletion of a functional domain of p53 [43 (link)] were a gift from Dr. Vogelstein. The HT-1080 p53 KO, HT-1080 p21 KO and SK-HEP1 p53 KO cell lines were genetically engineered using CRISPR technology [19 (link)]. All other cell lines were obtained from ATCC.