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. 2019 Aug 29;117(7):1352–1363. doi: 10.1016/j.bpj.2019.08.028

Figure 4.

Figure 4

ANAP spectral imaging afforded detection of hydrophobicity in intracellular environment. (a) A schematic of the inactivation gate depicting the location of the four sites tested, Q1475, F1485, E1488, and E1489is given. F1485 is a part of the IFM motif, known as the inactivation particle. Approximate location of the membrane is depicted by the dashed line. Structure adapted from Protein Data Bank: 5XSY (27). (b) ANAP spectra, laser line background-subtracted and normalized to the sum of intensities under each peak, are shown. ANAP at each of the four sites is compared with background ANAP control (wild-type Nav1.5+10 μM ANAP, black). Dashed box shows inset in which spectra are zoomed in on the peak. n = 12 cells for each condition, representative of two experiments, error bars are mean ± SE. (c) Fluorescence ratio F456 nm/F499 nm measured for each condition, error bars are mean ± SE. ∗∗∗∗p < 0.0001, ANOVA with Tukey’s multiple comparisons test. To see this figure in color, go online.