Figure S3.
Representative phasor plots A and B track data points recorded before and at 5, 10, and 15 minutes after water-soluble cholesterol (W S C) treatment. The horizontal axis represents the G factor, and the vertical axis represents the S factor, with each subplot displaying the fluorescence response through color-coded and differently shaped data points clustered along a universal semicircle. Section A focuses on Naive D R G neurons, showing a baseline E (F R E T) of 0.17 that remains stable at 0.18 throughout the treatment, while Section B shows S N I D R G (injured) neurons starting at a lower baseline of 0.05 and rising significantly to 0.13, 0.14, and 0.16 over the 15-minute interval. These annotated E (F R E T) values highlight changes in energy transfer over time, demonstrating that while naive neurons maintain stable molecular clustering, cholesterol enrichment restores the impaired membrane domain density in injured neurons. All values are approximated.
CTxB-based FLIM-FRET responses to cholesterol enrichment in naïve and SNI nociceptor DRG neurons. (A and B) Representative phasor plots showing membrane-localized fluorescence in naïve (A) and SNI (B) nociceptor DRG neurons labeled with CTxB AF-488 and CTxB AF-647, recorded before and at 5, 10, and 15 min after WSC treatment.