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. 2017 Nov 13;6:e29176. doi: 10.7554/eLife.29176

Figure 2. Ser57 phosphorylation of ubiquitin contributes to ppz mutant phenotypes.

Figure 2.

(A) Quantitative immunoblot analysis was performed on lysates from wild-type or ppz mutant yeast cells (SUB280 background) expressing either wild-type ubiquitin or Ser57Ala ubiquitin. (B) The results for (A) were quantified over multiple experiments (n = 3). (C) Quantitative immunoblot analysis was performed on lysates from wild-type or ppz mutant yeast cells (SUB280 background) expressing either wild-type ubiquitin or Ser57Ala FLAG-ubiquitin (driven by the TEF1 promoter) following addition of cycloheximide (CHX). (D and E) The results for (C) were quantified over multiple experiments (n = 3). (F) Wild-type and ppz mutant yeast strains (SUB280 background) expressing only wild-type or Ser57Ala ubiquitin were plated in 10-fold serial dilution on the indicated media. Plates were imaged after three days of growth at 26°C. In all panels, double asterisk (**) indicates p<0.005 and single asterisk (*) indicates p<0.05.

Figure 2—source data 1. This spreadsheet contains the quantification and statistical analysis for total ubiquitin levels (Figure 2B) and for ubiquitin degradation in a cycloheximide chase experiment (Figure 2D–E).
elife-29176-fig2-data1.xlsx (11.8KB, xlsx)
DOI: 10.7554/eLife.29176.011