Figure 3. Mitochondria function upstream of the GAAC response during autophagy. Cells were grown to log phase in SCD medium without uracil, then transferred to amino acid starvation medium with 2% glucose for 6 h. (A) Gcn2-dependent phosphorylation of eIF2α was determined by Western blot analysis using α-phospho Ser-51 eIF2α and α-Sui2 (eIF2α) antibodies. Quantification represents the ratio of phosphorylated eIF2α Ser-51 and total eIF2α (Sui2) signal with fold change relative to WT at t = 0 h. Data are presented as means ± SD of three independent experiments. (B) GCN4 de-repression was measured using the p180 GCN4 URE-1 to -4:lacZ reporter plasmid expressed in all strains. β-Galactosidase (β-gal) activity was determined as described in the legend to Fig. 2. Data are presented as means ± SD of three independent experiments.
Figure 3.

Mitochondria function upstream of the GAAC response during autophagy. Cells were grown to log phase in SCD medium without uracil, then transferred to amino acid starvation medium with 2% glucose for 6 h. (A) Gcn2-dependent phosphorylation of eIF2α was determined by Western blot analysis using α-phospho Ser-51 eIF2α and α-Sui2 (eIF2α) antibodies. Quantification represents the ratio of phosphorylated eIF2α Ser-51 and total eIF2α (Sui2) signal with fold change relative to WT at t = 0 h. Data are presented as means ± SD of three independent experiments. (B) GCN4 de-repression was measured using the p180 GCN4 URE-1 to -4:lacZ reporter plasmid expressed in all strains. β-Galactosidase (β-gal) activity was determined as described in the legend to Fig. 2. Data are presented as means ± SD of three independent experiments.

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