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. 2017 Sep 4;216(9):2691–2700. doi: 10.1083/jcb.201704056

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© 2017 Cruz-Garcia et al.

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Figure 1. — SOD1 and Acb1 are secreted by the same pathway in response to nutrient starvation. (A and B) Secretion of SOD1 is induced by nutrient starvation. Wild-type cells were grown in nutrient-rich medium to mid-logarithmic phase (growth). An aliquot of this culture was washed twice and incubated in 2% potassium acetate for 2.5 h (starvation). Cell wall proteins were extracted from equal numbers of growing and starved cells followed by precipitation with TCA. Lysates and cell wall–extracted proteins (wall) were analyzed by Western blotting. SOD1 levels were quantified as the ratio of wall to lysate SOD1 and are represented as percentages of the starved wild-type culture. (C and D) Grh1 is required for SOD1 secretion. The ratio of wall to lysate SOD1 in wild-type and grh1Δ cells incubated in 2% potassium acetate for 2.5 h was determined as in A and B. (E and F) SOD1 secretion requires a subset of ESCRT proteins. The ratio of wall to lysate SOD1 in wild-type, vps23Δ, snf7Δ, and vps4Δ cells incubated in 2% potassium acetate for 2.5 h was determined as in A and B. (165X) wall indicates that the wall sample represents the material from 165-fold the number of cells analyzed in the lysate sample. Results in B, D, and F are shown as the means ± SEM of three experiments.