MMTV-PyMT-S2WTP3 cells were infected with lentiviral particles expressing the pHAGE-deltaOA-zsGreen plasmid as previously described6 (link), and infected cells were isolated by FACS.
MDA-MB-453 and BT474 cells were transient transfected with pBabepuro3-p16Flag (Addgene Cat # 24934) using Lipofectamine 3000 (Thermo Fisher) according to the manufacturer’s instructions. 72 hours after transfection cells were selected in puromycin for 48 hours. p16 overexpression was confirmed by anti-FLAG western blots.
For overexpression of DNMT1, MDA-MB-453 cells were transfected with the pcDNA3/Myc-DNMT1 vector (Addgene Cat #36939) using Lipofectamine 3000 according to the manufacturer’s instructions. Cells were treated with abemaciclib 72 hrs after transfection.
MDA-MB-453 and BT474 cells were transient transfected with pBabepuro3-p16Flag (Addgene Cat # 24934) using Lipofectamine 3000 (Thermo Fisher) according to the manufacturer’s instructions. 72 hours after transfection cells were selected in puromycin for 48 hours. p16 overexpression was confirmed by anti-FLAG western blots.
For overexpression of DNMT1, MDA-MB-453 cells were transfected with the pcDNA3/Myc-DNMT1 vector (Addgene Cat #36939) using Lipofectamine 3000 according to the manufacturer’s instructions. Cells were treated with abemaciclib 72 hrs after transfection.
