Figure 3. The involvement of ESCRTs in CUPS formation, stability and disassembly.
(A) Wild type and the indicated deletion strains expressing Grh1-2xGFP were grown to mid-logarithmic phase, washed twice, and incubated in 2% potassium acetate for 30 min and 2.5 hr to assess CUPS biogenesis and stability, respectively. Cells were grouped in 3 classes: CUPS (1–3 large punctae per cell), intermediate (large and small punctae), no CUPS (multiple small punctae). Between 50–200 cells were counted for each strain/condition from 3 independent experiments. (B) Effect of loss of Snf7 of ESCRT-III at 30 min and 2.5 hr starvation. (C) Wild type and vps4Δ cells were starved for 2.5 hr as in (A) to allow CUPS formation. Cells were washed once and cultured in rich media for 45 min. Scale bars = 2 μm.