To begin the cell culture, prepare the DMEM culture medium and solutions required for cell culture. Seed 1 x 10 to the 6th HEK293 cells in a 60-millimeter plate or a 6-well plate per treatment condition plus control. The next day, treat the cells with DNA-protein crosslinks, or DPCs, inducers of choice.
To induce DPCs, their ubiquitylation and SUMOylation, collect the cells at 20, 60, and 180 minutes, or two hours in case of non-enzymatic DPCs. To induce the PARylation of TOP1-DPCs, pre-treat the cells with 10 micromolar poly-ADP-ribose glycohydrolase, or PARG inhibitor PDD00017273, for one hour before co-treating with 20 micromolar camptothecin for 20, 60, and 180 minutes.
Sun, Y. Cell Culture and Drug Treatment in the Human Embryonic Kidney 293 (HEK293) Cell Line. J. Vis. Exp. (Pending Publication), e200208, doi:10.3791/200208 (2023).