Figure 5.
Rad26 co-localizes with transcription upon UV irradiation. ChIP experiment. Triple HA-tagged Rad26 was immunoprecipitated from indicated strain using 12CA5 antibodies. (A) Rad26 localizes to the ACT1 promoter after UV. Chromatin extracts were prepared from cells either unirradiated or from irradiated cells at indicated time points (min) post 70 J/m2 of UV. DNA was amplified from precipitates using primers corresponding to the promoter and CDS of the ACT1 gene. Quantification of the PCR product signals are displayed in histograms below corresponding PCR series as in Figure 2. (B) Rad26 cross-links to RNAP II after UV. Rad26 was immunoprecipitated from chromatin extract prepared from cross-linked cells either unirradiated or from cells irradiated with indicated doses (in J/m2) at indicated time points (minutes) post UV. Precipitates were run across a SDS–PA gel blotted to a PVDF membrane and RNAP II was detected using H14 antibody.