RNA polymerase IIo ChIP-seq by employing the K1268R RPBI mutant and TC-NER deficient HeLa cells

RNA polymerase IIo ChIP-seq was performed to investigate the role of ubiquitination at the K1268 residue of human RPB1: the K1268R mutant lost transcription-coupled nucleotide excision repair (TC-NER) ability, resulting in induction of premature aging phenotypes. Wild type, the K1268R mutant, and TC-NER deficient HeLa cells were employed for the experiment. RPB1 molecules were immunoprecipitated with anti- phopho-Ser2-CTD (RPB1-Ser2: 3E10, Ab5095), or -Ser5-CTD (RPB1-Ser5: 3E8) antibodies, as well as with an anti-pan-RNA pol II (RPB1-pan: A304405A) antibody.