PMPs survive STZ cytotoxicity and produce larger colonies with a higher yield of insulin+ progeny. PMPs survive STZ cytotoxicity and produce larger colonies.

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PMPs survive STZ cytotoxicity and produce larger colonies with a higher yield of insulin+ progeny. PMPs survive STZ cytotoxicity and produce larger colonies with a higher yield of insulin+ progeny. A: Log2 fold expression change (at P < 0.05) of genes involved in β-cell development pathway in insulin+Glut2low vs. insulin+Glut2high cells. B: Results of glucose tolerance tests performed on day 4 after STZ injection. Repeated-measures ANOVA comparing the two groups showed a significant effect of STZ/vehicle treatment (F[1,8] = 6.04, P = 0.045, n = 5 mice/group). C: Scatterplot of measured FPKM (fragments per kilobase of transcript per million fragments) for insulin+Glut2high and insulin+Glut2low cells from control mice (CTRL). Only genes with FPKM >0 in both samples are shown. D: Differential gene expression in insulin+Glut2low cells of STZ-treated mice in comparison with control mice. Cell cycle genes, defined based on REACTOME annotations, are shown in E, with red bars corresponding to the “leading-edge” set of cell cycle genes. Cell cycle genes are significantly upregulated in insulin+Glut2low cells of STZ-treated mice relative to control mice (Wilcoxon rank sum test, P < 0.002). E: Expression of the leading-edge set of cell cycle genes across different samples. The majority of these genes are specifically expressed in Glut2low cells of STZ-treated mice. F: Frequencies and diameter measures of PMP spheres from STZ-induced diabetic and control islets. Two-way ANOVA revealed a significant interaction of STZ-induced diabetes/vehicle treatment and the diameter of spheres (F[1,11] = 22.5, P = 0.006), with a main effect of STZ/vehicle treatment (F[1,8] = 6.04, P = 0.045). Bonferroni posttest showed a significant increase of large spheres in the STZ-treated group (P < 0.05, n = 5 mice/group). G: Immunofluorescent images of clonal differentiated PMP colonies derived from diabetic and control pancreata showing the insulin-expressing cells (red). Scale bar, 20 μm. H: Frequencies of insulin+ and glucagon+ progeny produced by differentiated clonal PMP spheres from STZ-induced diabetic islets and controls. Two-way ANOVA indicated the significant interaction of diabetes and hormone+ cell production (F[1,28] = 7.19, P = 0.0121), with Bonferroni posttest showing increased number of insulin+ cells in differentiated diabetic PMP colonies (P < 0.05, n = 20 spheres/group). Changes in glucagon+ cell production were nonsignificant. I: FPKM of genes involved in β-cell development pathway are compared between insulin+Glut2high and insulin+Glut2low cells from diabetic and control islets. Values are means ± SEMs where error bars are shown. FDR, false discovery rate. Rozita Razavi et al. Diabetes 2015;64:1311-1323 ©2015 by American Diabetes Association