Objective to explore the mechanism of STAT3 regulating PD-L1 expression at transcriptional level after LIFR overexpression in pancreatic cancer cells.

The assays were performed in the standard manner to determine the influence of LIFR on the capacity of STAT3 (Cell signaling Technologyv, 124H6, cat#: 9139, 1µg) binding to DNA. Specific DNA fragments were obtained, purified, and subjected to PCR amplification using a NovoNGS® CUT&Tag 2.0 High-Sensitivity Kit (Novoprotein Scientific Inc), according to the protocol of the manufacturer. Berry Genomics Corporation (Beijing, China) was trusted to prepare the libraries and perform the sequencing. Overall design: To uncover the target genes directly regulated by STAT3 that is activated by LIFR overexpression in Mia PaCa2 cells. To this end, we performed Cleavage Under Targets and Tagmentation (CUT&Tag) sequencing with anti-STAT3 antibody to probe the enhanced genomic binding sites of STAT3 upon LIFR activation