Fig. 7

Characterization of S5D3- S7D10 (later-stage) clusters. A Dithizone staining of late-stage aggregates and various donor human islets, scale bar = 500 µm B gene expression profile of later-stage cells relative to donor human islets, scale bar = 500 µm (n = 13–24 biological replicates; mixed-effects analysis with Tukey post-test, MAFA, MAFB, UNC3: paired two-tailed t-test). C Flow cytometry of S6D7 aggregates with the indicated markers. D Representative INS, GCG, SST immunohistochemistry staining of S6D7, S7D8 aggregates and human islets, scale bar = 250 µm. E Schematic of perifusion protocol. F Dynamic human insulin secretion and fold change in response to various secretagogues (n = 15 stage 6 biological replicates, n = 8 biological replicates stage 7, 10 donor human islet preps; line = mean, shaded area = SEM; numbers above the whisker in fold change plots indicate the mean). All box and whisker plots show individual points with median and interquartile range; bar charts show individual points with mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. S5D3 = stage 5 day 3, S6D7 = stage 6 day 7, S7D8 = stage 7 day 8