Fig 1: Integrin activation mediates ß-cell orientation and glucose-dependent Ca2+ responses.(A) Immunofluorescence staining of phospho-FAK, E-cadherin, and insulin showed that isolated ß cells, cultured on bovine serum albumin (BSA)-coated coverslips, were disorganised. Cells were multilayered and the phospho-FAK staining scattered at the edges of the footprint of the cells, also see orthogonal sections. (B) In contrast cells cultured on laminin-coated coverslips showed extensive, punctate phospho-FAK located at the cell footprint (as shown in the orthogonal section) and organised E-cadherin staining at the cell junctions. Immunofluorescence staining of isolated ß cells (insulin; blue), grown on BSA- (C) or laminin- (D) coated coverslips showed enriched ELKS (green) and liprin (red) staining at the laminin–cell, but not BSA–cell interface, compared with the cytoplasm, see orthogonal sections (XZ) for cells cultured on BSA (E) or laminin (G). (F) Average fluorescence intensity of both ELKS (Student’s t-test, p < 0.001) and liprin (Student’s t-test, p < 0.05) were significantly lower at the BSA–cell interface compared with the cytosol (36 regions of interest [ROIs], n = 6 cells from three animals). (H) In the cells cultured on laminin the average fluorescence intensity of ELKS and liprin were significantly higher at the laminin–cell interface compared with the cytosol (Student’s t-tests, p < 0.001) (36 ROIs, n = 6 cells from three animals). Using Fura-2-loaded, isolated ß cells cultured on BSA, high glucose induced a modest, short-lasting response (I) that contrasted with the large response and sustained oscillations when the cells were cultured on laminin (J), with a significant reduction in area under the curve (AUC) of the response (K, Student’s t-test p < 0.001, n = 36 cells on laminin and n = 21 cells on BSA). Scale bars in (A, B) 10 µm, all others 5 µm. *** shows statistical significance at p<0.001. Figure 8—source data 1.Scaffold protein distribution analysis.