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. 2014 Aug 12;42(16):10503–10515. doi: 10.1093/nar/gku731

Figure 3.

Figure 3.

The proteasome associates with RNAPII and transcribed genes. (A) RNAPII was purified in the presence of DUB and proteasome inhibitors using an RPB3-TAP strain and levels of Rpb1 and co-purifying proteasome detected with 8WG16 and an anti-20S core antibodies, respectively (upper panel). Columns and error bars represent the mean ± standard deviation from four independent experiments (bottom panel). (B–D) To assess recruitment of the proteasome to actively transcribed genes, ChIP experiments were performed with a RPB3-TAP and a PRE1-TAP strain. Enrichment of Rpb3 (B), Pre1 (C) and Pre1 relative to Rpb3 (RNAPII) (D) at the genes ADH1, ACT1 and PMA1 was quantified by real-time PCR with the indicated primers (upper panel). (E–G) Experiment as in B–D after inhibition of transcription by treatment with 250 μg/ml 6AU for 2.5 h. Columns and error bars represent the mean ± standard deviation from three independent experiments (bottom panels). P-values are listed in Supplementary Table S3.