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. 2016 Sep 19;17(11):1590–1608. doi: 10.15252/embr.201642209

Figure 8. Effect of chlorpromazine on fusion activity and morphology of double‐tagged vacuoles.

Figure 8

  1. Yeast strain expressing a plasmid with Vph1‐GFP was grown in SC‐URA medium for 16 h at 30°C. The mobility of Vph1‐GFP in the absence (upper panels) or presence (lower panel) of CPZ (150 μM) was assessed by FRAP.
  2. The histogram shows the GFP signals in the bleached areas during the FRAP procedure. Means ± s.d. are shown for 20 cells of three independent experiments.
  3. The half‐time is shown. Means and s.d. are shown for 20 cells of three independent experiments.
  4. Yeast cells were grown in SC medium for 16 h at 30°C. After staining with FM4‐64, cells were treated with CPZ (150 μM). Vacuole morphology was assessed by confocal microscopy before and after 5 and 20 min of CPZ addition. Scale bar: 5 μm.
  5. Fusion activity: Vacuoles were isolated from BJ cells carrying NyV1‐S9‐EGFP and DKY cells deleted for Nyv1 and carrying VaM3‐S9‐mCitrine or from wild‐type cells and incubated in standard fusion reactions in the presence or absence of chlorpromazine (CPZ, 150 μM) and anti‐Vam3 (3 μg). Content mixing was assayed via alkaline phosphatase activity. Means and s.d. are shown for three independent experiments.