Figure 3.
Figure 3. The Snx4 complex mediates Atg27 vacuole-to-endosome retrograde transport. (A) Schematic predicting Atg27 accumulation on the vacuole membrane in retromer and recycling double mutants. (B) Atg27-GFP localization in vps35Δ snx4Δ cells after 1 h of rapamycin treatment. (C) The percentage of cells with Atg27-GFP on the vacuole. (D) Atg27-GFP localization in vps35ts snx4Δ mutants after shift to 37°C with rapamycin treatment for 1 h. (E) Quantification of Atg27-GFP fluorescence intensity on the vacuole from D and Fig. S2 C. (F) Cells were treated with rapamycin for 1 h, Atg27-GFP was immunoprecipitated, and interacting Snx4-3XHA was detected by immunoblot. For all quantification shown in this figure, at least 100 cells were measured, and the data from three independent experiments were used for statistical analysis; two-tailed Student’s t test. Error bars represent SD. IP, immunoprecipitation; n.s., not significant. Bar, 2 µm.

The Snx4 complex mediates Atg27 vacuole-to-endosome retrograde transport. (A) Schematic predicting Atg27 accumulation on the vacuole membrane in retromer and recycling double mutants. (B) Atg27-GFP localization in vps35Δ snx4Δ cells after 1 h of rapamycin treatment. (C) The percentage of cells with Atg27-GFP on the vacuole. (D) Atg27-GFP localization in vps35ts snx4Δ mutants after shift to 37°C with rapamycin treatment for 1 h. (E) Quantification of Atg27-GFP fluorescence intensity on the vacuole from D and Fig. S2 C. (F) Cells were treated with rapamycin for 1 h, Atg27-GFP was immunoprecipitated, and interacting Snx4-3XHA was detected by immunoblot. For all quantification shown in this figure, at least 100 cells were measured, and the data from three independent experiments were used for statistical analysis; two-tailed Student’s t test. Error bars represent SD. IP, immunoprecipitation; n.s., not significant. Bar, 2 µm.

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