Figure 5

Primary murine islet beta-cells cultured on laminin matrix show increased levels of FGFR5 protein and glycolytic enzyme activity. To examine the effects of laminin induced transcript changes in a more physiologically relevant model, primary murine islets were dispersed on laminin matrix and the protein levels of previously tested genes were measured. Islets were dispersed onto poly-D-lysine (PDL) or laminin matrix (LM) in glass bottom dishes and transduced with mChe or R5 under a rat insulin promoter to identify beta-cells. (a) Endogenous FGFR5 immunofluorescence intensity of beta-cells. (b) Endogenous PKM2 immunofluorescence intensity of beta-cells plated on PDL or LM, or beta-cells overexpressing the R5 construct (R5) plated on PDL. (c) The Apollo-NADP⁺ response to glucose (1 to 15 mM) of beta-cells plated on PDL and LM reported as the change in anisotropy (ΔAnisotropy). (d) Dispersed beta-cells plated on PDL, plated on PDL and expressing the R5 construct (R5), or plated on LM were incubated with 15 mM glucose and 2 µM 2-NBDG for 20 min at 37 °C prior to imaging. Bars show the average indicated value ± SEM; symbols represent individual responses of n = 3 independent experiments. *, p < 0.05 using ANOVA and one-tailed t-test.