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. 2022 Apr 19;23(6):e53835. doi: 10.15252/embr.202153835

Figure 2. Gid4 and Gid10 compete for binding to GIDAnt .

Figure 2

  1. Lysates from wildtype, ΔGid4, and Gid4ΔC strains expressing endogenously tagged 3xFLAG‐Gid10 that were grown at 42°C for 1 h, and then returned to 30°C for the indicated timepoints were run on an SDS‐PAGE gel and immunoblotted with αFLAG and αPGK.
  2. Wildtype and Gid10 overexpressing yeast strains expressing endogenously tagged Gid4 were grown for 19 h in YPE, and transitioned to YPD for the indicated timepoints. Lysates were run on an SDS‐PAGE gel and immunoblotted with αFLAG and αPGK. Points represent mean, error bars represent standard deviation (n > 3 biological replicates).
  3. Fbp1 protein abundance in wildtype and Gid10 overexpressing yeast strains grown in YPE for 19 h, and following 30 min and 2 h recovery growth in YPD. Bars represent mean, error bars represent standard deviation, significance was determined by one‐way ANOVA (unpaired) followed by Tukey’s multiple comparison test, * indicates P < 0.05, ** indicates P < 0.01, (n = 3 biological replicates).
  4. In vitro competition assay probing the ability of N‐terminally 2xStrep‐tagged Gid4 (2xS‐Gid4) to exchange with Gid10 in the GIDSR10 complex. Proteins after Strep‐Tactin pull‐down were visualized by Coomassie‐stained SDS‐PAGE.