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Pol II-Med and Pol II-TFIIF complexes are dynamic in nuclear extract. Purified Pol II-Med or Pol II-TFIIF was incubated with nuclear extract from an Rpb11-HA-tagged strain under transcription conditions without DNA template and in the presence or absence of NTPs as indicated. Reactions were incubated at 25°C for 20 min and immune precipitated with HA antibody. Precipitated proteins were analyzed by SDS-PAGE and Western blotting with Flag M2 and anti-HA antibodies.
