Exploratory experiment designed to discover new patterns targeting IRF1, NLRC5, CIITA in cultured cells with reporter constructs. Primary outcome: IRF1 transcriptional activity on Nlrc5 and Ciita promoters
Dual-luciferase reporter assays and chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) to determine how NPM1 regulates IRF1's DNA binding activity to the Nlrc5 and Ciita promoters. These experiments revealed the mechanism by which NPM1 sequesters IRF1 from binding to target promoters, thereby suppressing IRF1-mediated expression of MHC-I and MHC-II molecules in tumor cells. The study demonstrated that NPM1 associates with IRF1 and prevents its transcriptional activity on key antigen presentation genes.
Dual-luciferase reporter assays to measure promoter activity and ChIP-qPCR to assess IRF1 DNA binding to target promoters
NPM1 would suppress IRF1-mediated transcriptional activation of MHC genes
Reduced IRF1 binding and transcriptional activity in presence of NPM1
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