CUT&Tag analysis of H3K4me3, H3K27Ac,H2A119Ub and H3K27me3 in treated KPC1199 murine PDAC cell line

To gain mechanistic insight into how Epigenetic factors reprogramming tumor immune microenvironment in response to IFN? stimulation. we performed CUT&Tag sequencing in murine PDAC cells.For comparison of the transcription profile between IFN?-PD-L1all and IFN?+PD-L1all cells, we treated KPC1199 cells with vehicle control or 10 ng/mL recombinant mouse IFN? for 20 hours and harvested them. For comparison of PD-L1hi with PD-L1all, cells were either sorted with the top 15% PD-L1+ or total of IFN?+PD-L1all PDAC cells, and freshly collected as soon as possible. Hyperactive pA-Tn5 Transposase for CUT&Tag kit from Vazyme (TD901), and antibodies against H3K4me3 from Abcam (ab213224), H3K27Ac from Abcam (ab4729),H2A119Ub from Cell Signaling (8240T) and H3K27me3 from Abcam (ab6002) were employed. Trueprep index kit v2 and v3 for illumina were used to establish DNA library. Overall design: For library preparation, murine PDAC cell lines withIFN?-PD-L1all, IFN?+PD-L1all and IFN?+PD-L1hi were first established. CUT&Tag and DNA library were prepared according to manufacturer's instructions.