Figure S2.
Confocal imaging shows the effects of cholesterol extraction or supplementation on membrane localization of G R A M-W-e G F P. In panel A, t s A cells overexpressing G R A M-W-e G F P are shown before and after 5 minutes of treatment with 2.5 millimeters beta-cyclodextrin (beta-C D). Line scans of an area indicated by white dashed lines highlight the increase in cytosolic green fluorescent protein (G F P) fluorescence after beta-C D treatment. In panel B, t s A cells overexpressing G R A M-W-e G F P are shown before and after 15 minutes of treatment with 0.5 milligrams per milliliter water-soluble cholesterol (W S C), highlighting the increase in membrane-localized G F P fluorescence after cholesterol supplementation. The horizontal axis of the graphs represents distance in micrometers, and the vertical axis represents fluorescence in arbitrary units. All values are approximate.
Confocal imaging showing the effects of cholesterol extraction or supplementation on membrane localization of GRAM-W-eGFP. (A) tsA cells overexpressing GRAM-W-eGFP, before and after 5 min of treatment with 2.5 mM β-CD. Line scans of an area such as that indicated by white dashed lines are plotted to highlight the increase in cytosolic GFP fluorescence after β-CD. (B) tsA cells overexpressing GRAM-W-eGFP, before and after 15 min of treatment with 0.5 mg/ml WSC, highlighting the increase in membrane-localized GFP fluorescence after cholesterol supplementation.