Interaction factors of DDB2 immunopurified from UV irradiated cells

HEK293F cells expressing FLAG-tagged DDB2 were exposed with UV-C. After 15 min incubation, cell lysate was prepared and chromatin fraction was fractionated. The resulting chromatin sample was digested by micrococcal nuclease treatment. The cellular UV-DDB (DDB1-DDB2 complex) bound to chromatin was then immunopurified by agarose beads conjugated with the anti-FLAG M2 antibody. After DDB2 immunopurification, elute fraction was subjected to SDS-PAGE and stained by CBB. Target proteins in the excised gels were in-gel digested with trypsin and identified by nano LC-MS/MS analysis.