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. Author manuscript; available in PMC: 2015 Jan 9.
Published in final edited form as: Mol Cell. 2014 Jan 2;53(1):148–161. doi: 10.1016/j.molcel.2013.12.003

Figure 3. Stability of Grr1 candidate substrates.

Figure 3

(A) Epitope-tagged candidate Grr1 substrates were expressed in GRR1 or grr1Δ cells. To rescue slow growth, all grr1Δ cells are also rgt1Δ. Asynchronous cultures were treated with cycloheximide (CHX) for the indicated number of minutes and anti-Myc western blots were performed on whole cell extracts. Western blot of Cdc28 is shown as a loading control. (B) A subset of candidate substrates were analyzed during the cell cycle by Western blots of whole cell extracts. Cells were released from α-factor-mediated G1 arrest and collected at different time points. α-factor was again added after 60 minutes to re-arrest cells in the subsequent G1. Clb2 and Cdc28 are shown to monitor cell cycle progression and as a loading control, respectively. Asynchronous cells are shown for comparison.