Figure 7

FGFR5 overexpression in primary human beta-cells induces a more robust glucose stimulated NADPH response​. To further support the physiological relevancy of our findings, R5 overexpression was examined in primary human islet samples. Human islets were dispersed to single cells on PDL treated glass bottom dishes and co-transduced with Apollo-NADP⁺ sensor (CMV promoter) and either mChe, R5, or R5ΔC (RIP promoter). (a) Widefield fluorescence (WD Fluor) and anisotropy images from a representative beta-cell at multiple glucose concentrations, as indicated. (Scale bar = 5 μm; anisotropy lookup table for widefield anisotropy =  × 10^–2). (b) Apollo-NADP⁺ response (Anisotropy) of mCherry positive beta-cell cells overexpressing R5, mChe, and R5ΔC in response to sequential glucose concentrations, as indicated. (c) The Apollo-NADP⁺ responses of mChe, R5 and R5ΔC expressing beta-cells (traces shown in Fig. S9) were classified as either strong (orange) or weak (dark blue) glucose responders based on the mean mChe cell response. Bars indicate % cell distribution for each construct and represent the average ± SEM of n = 7 independent experiments. *, p < 0.05 using ANOVA followed by one-tailed t-test.