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. 2011 Dec 12;195(6):965–978. doi: 10.1083/jcb.201104062

Figure 5.

Figure 5.

Osh4p delivers DHE to PI(4)P-containing liposomes. (A) Osh4p loaded with DHE (∼0.5 µM Osh4p[DHE]) was incubated at 30°C in buffer. At the indicated time, 30 µl of a stock suspension of liposomes (5 mM lipids) was injected (final concentration = 240 µM). The release of DHE was followed by measuring the increase in Osh4p fluorescence at 340 nm (λex = 285 nm). The liposomes contained DOPC and, when indicated, 2 mol% of DOPS, PI, PI(3)P, PI(4)P, PI(5)P, or PI(4,5)P2. (B) DNS-Osh4p loaded with DHE (∼0.5 µM) was mixed with 240 µM liposomes with (left) or without (right) 5 mol% ergosterol at 30°C. The release of DHE was followed by measuring the diminution in FRET between DHE and DNS (λex = 310 nm; λem = 510 nm). In all assays, the recordings were corrected for the light-scattering signal and the dilution effect was due to liposome addition.