The targeted genes of KIF11 of PLC8024 cell after drug treatment
We found that KIF11 played an inportant role in PLC8024 apoptosis after drug treatment Overall design: The CUT&Tag assay was performed in PLC8024 cells using the NovoNGS® CUT&Tag® 4.0 High-Sensitivity Kit (designed for Illumina®) (Novoprotein, N259-YH01, Shanghai, China), following the manufacturer's instructions. The experiment included four groups: an IgG negative control group, a Pol II positive control group, a PLC8024 control group, and a PLC8024 group treated with drug(10 nM for 6 hours).The key experimental steps involved ConA magnetic bead purification, cell preparation, binding of nuclei to ConA magnetic beads, overnight incubation with a primary antibody, secondary antibody binding followed by washing, transposition complex binding and washing, a one-hour fragmentation reaction, termination of fragmentation, DNA extraction, PCR amplification, purification of PCR products, library construction using index primers, and sequencing. Finally, specific primers were designed for qPCR analysis based on the sequencing results.