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Figure 2 | BMC Molecular Biology

Figure 2

From: Rrd1 isomerizes RNA polymerase II in response to rapamycin

Figure 2

Analysis of the GST-CTD and its interaction with Rrd1. A) Comparison of the expression and phosphorylation status of the GST-CTD between parent and rrd1 Δ mutant cells following rapamycin exposure. The indicated cells expressing GST-CTD were treated with (+) and without (-) rapamycin (200 ng/ml for 30 min) and total protein extracts were probed for Ser-2 phosphorylation (H5) or Ser-5 phosphorylation (H14). The membranes were stripped and reprobed with anti-GST antibody. B-D) Retention of Rrd1-MYC by GST-CTD affinity beads. B) The input (5% of the total amount of protein extracts added to the beads) of parent cells expressing Yap8-MYC and Rrd1-MYC from the endogenous locus. Western blot analysis was done using anti-MYC antibody. C) and D) Total protein extracts derived from the parent or parent expressing either Yap8-MYC or Rrd1-MYC were incubated with the empty beads or beads containing either GST-CTD or GST-Apn1 (see Methods). The beads were then washed and an aliquot examined for retention of the MYC tagged proteins using anti-GST antibodies (C) or anti-GST antibodies (D). Results shown are representative of two independent experiments.

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