Figure 3.
Figure 3. Membrane ordering in the vacuolar domains. (A) Intact vacuoles from cells expressing Vph1-GFP were incubated with Fast DiI for 30 min and imaged by fluorescence microscopy. (B) Images of live cells expressing Vph1-GFP and Ivy1-mCherry (shown in Fig. 1 C). (C) Images of isolated vacuoles from cells expressing Vph1-GFP and strained with filipin. In B and C, arrows indicate inward bending of sterol-enriched domains. (D) Generalized polarization (GP) of liposomes (left) and cellular membranes (right) incubated with Laurdan for 1 h at 30°C (means ± SD, n = 4–5 independent experiments). chol, cholesterol; DOPC, dioleoylphosphatidylcholine; SM, sphingomyelin; vac, vacuole. Where indicated, a two-tailed t test was used to calculate p-value. Bars: (A) 2 µm; (B and C) 1 µm.

Membrane ordering in the vacuolar domains. (A) Intact vacuoles from cells expressing Vph1-GFP were incubated with Fast DiI for 30 min and imaged by fluorescence microscopy. (B) Images of live cells expressing Vph1-GFP and Ivy1-mCherry (shown in Fig. 1 C). (C) Images of isolated vacuoles from cells expressing Vph1-GFP and strained with filipin. In B and C, arrows indicate inward bending of sterol-enriched domains. (D) Generalized polarization (GP) of liposomes (left) and cellular membranes (right) incubated with Laurdan for 1 h at 30°C (means ± SD, n = 4–5 independent experiments). chol, cholesterol; DOPC, dioleoylphosphatidylcholine; SM, sphingomyelin; vac, vacuole. Where indicated, a two-tailed t test was used to calculate p-value. Bars: (A) 2 µm; (B and C) 1 µm.

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