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. 2024 Feb 28;128(9):2154–2167. doi: 10.1021/acs.jpcb.3c07006

Figure 1.

Figure 1

Characterization of Flipper, NR12S, and NR12A lifetimes in different lipid environments. Spectral fluorescence lifetime measurements of the probes in LUVs were carried out within 500–700 nm in intervals of 20 nm. Multiexponential curve fitting was performed for the fluorescence decays. (A) Normalized intensity spectra of Flipper (left), NR12S (middle), and NR12A (right) in varying lipid environments. (B) Normalized fluorescence decays (full spectrum, 500–700 nm) of Flipper (left), NR12S (middle), and NR12A (right) in Δ9cis DOPC or DPPC:Chol 50:50. (C) Normalized fluorescence decays at 570 nm vs 650 nm of Flipper (left), NR12S (middle), and NR12A (right) in POPC:Chol 80:20. (D) Spectrally resolved intensity-weighted lifetime of Flipper, NR12S, and NR12A in Δ9cis DOPC and the control AF488 in water. The line corresponds to the median of individual biological replicates (n ≥ 3). The band corresponds to the standard deviation. (E) Spectrally resolved intensity-weighted lifetime of Flipper (left), NR12S (middle), and NR12A (right) in different lipid environments with varying saturation indices. The line corresponds to the median of individual biological replicates shown with different symbols (n ≥ 3). The band corresponds to the standard deviation.