Fig. 1.
Snf7 binds the N-terminus of Doa4. (A) Schematic diagram of Doa4 and Snf7. (B) Affinity isolation of GST-Doa4 fusion proteins incubated with the indicated ESCRT-III subunits. The asterisk indicates GST-Doa41-80, which was detected because the anti-Snf7 antiserum was raised against GST-tagged Snf7. (C) Affinity isolation of GST-Doa41-348 incubated with Snf7, Snf7ΔMIM2 (L199D, V202D), Snf7ΔMIM1 (L231A, L234A) or Snf7Δα6 (Δ226-240). (D) Live-cell BiFC fluorescence microscopy images showing the interaction of Doa4-VN or Doa41-348-VN (light gray bar) with Snf7-VC. Vacuolar membranes are stained with FM4-64. Dashed lines indicate cell outlines. Scale bar: 4 μm. (E) Quantification of the mean fluorescence per cell based on ≥3 independent experiments. Error bars represent the ±s.e.m.; *P≤0.05, ***P≤0.001. n.s., not significant.