The colon cell line SW48 with heterozygous knock-in of BRAF V600E mutation and the parental SW48 cells were purchased from Horizon Discovery (#HD 103-003, Cambridge, UK). The heterozygous knock-in of BRAF-activating mutation (V600E) was generated by rAAV technology through homologous recombination and Cre recombinase of the Neo cassette. One allele was knocked in with BRAF V600E. It was verified that there was no selection cassette at the engineered locus. BCPAP, K1, OCUT1, TPC1, C643, KAT18, WRO, HTORI-3, M14, and the BRAF V600E knock-in and the parental SW48 cells were grown at 37 °C in RPMI-1640 medium with 10% fetal bovine serum (FBS, #F2442; Sigma-Aldrich, St Louis, MO). FB1, A375, CHL-1, and 293T cells were grown at Dulbecco's Modified Eagle Medium (DMEM) medium with 10% FBS. SK-MEL-1, SK-MEL-2, MeWo, and RKO cells were grown in EMEM medium with 10% FBS. The SK-MEL-3 cell was grown in McCoy’s 5A medium with 15% FBS. The TERT promoter region was amplified by PCR using primers 5′-AGTGGATTCGCGGGCACAGA-3′ (forward) and 5′-CAGCGCTGCCTGAAACTC-3′ (reverse)6; the BRAF V600E mutation hot spot region was amplified using primers 5′-TCATAATGCTTGCTCTGATAGGA-3′ (forward) and 5′-GGCCAAAAATTTAATCAGTGGA-3′ (reverse)50. The PCR products were subjected to Sanger sequencing for the detection of BRAF V600E and TERT promoter mutations. The K1 cell line is reported to be contaminated with the GLAG-66 cell line in the International Cell Line Authentication Committee (ICLAC) database. These two cell lines are both human PTC-derived. Our genetic analysis of the K1 cell line used in the present study confirmed typical heterozygous BRAF V600E and TERT C228T mutations. Cells were authenticated by short tandem repeat analyses and tested for mycoplasma. The K1 cell used here met the purpose of the present study to investigate the role of BRAF V600E and TERT promoter mutations in human cancer.