Extended Data Figure 2: Diabetes recovery in pre-pubertal mice.
From: Diabetes recovery by age-dependent conversion of pancreatic δ-cells into insulin producers
a, Evolution of glycemia (AuC) between 2.5 and 4 mpa, in pups and adults (see Fig. 1b) (Welch’s test, P = 0.0188). b, qPCR of insulin 2 messenger RNA after β-cell ablation; insulin 2 transcripts are 25-fold more abundant in pups than in adults at 2 mpa (n = 3 mice per group, each individual sample was run in triplicate in each reaction for a total of three independent reactions). Built-in Welch’s test (P = 0.0134, 0.0049). Error bars show s.d. c, Glucose tolerance tests (IPGTT) for DT-treated (4.5 mpa, n = 4) and age-matched controls (n = 4); note the fold increase between glucose injection and the glycaemic peak during IPGTT for each animal, and fold decrease between glycaemic peak and T120 (two-tailed unpaired t-test, PI = 0.5836, PII = 0.4937). d, Plasma insulin at time point (in min) T0, T15 and T30 during the IPGTT. Control: n = 4; DT: n = 4; two-tailed paired t-test (P = 0.0008). e, Insulin tolerance tests (ITT) performed 1.5 years after β-cell ablation at 2 weeks of age. Controls: n = 7; DT: n = 10. f, 4.5 months after β-cell ablation (at 2 weeks), three mice became normoglycaemic and received a second treatment with DT. Ablation of regenerated insulin+ cells in recovered mice leads to the appearance of glucagon+/insulin+ cells, corresponding to the type of ‘α-cell-dependent’ regeneration observed in adults (31% of insulin+ cells also contained glucagon; Supplementary Table 8). Arrow indicates glucagon+/insulin+ bihormonal cell. Error bars show standard error of the mean (s.e.m.). g, β-cell proliferation is very low in regenerating pups (Supplementary Table 9). Control: n1-month-old = 3, 6,006 insulin+-cells scored; n2-month-old = 3, 6,358 insulin+-cells scored; DT: n0.5 mpa = 5, 412 insulin+-cells scored; n1.5 mpa = 3, 675 insulin+-cells scored; Welch’s test (P = 0.1197, P = 0.0688). Error bars show s.e.m. h, Islet cell proliferation is increased (3.5-fold; Ki67+ cells) in islets of DT-treated pups at 0.5 mpa. Control: n1-month-old = 3, 95 islets scored; n1.5-month-old = 3, 94 islets scored; n2-month-old = 3, 90 islets scored; n2.5-month-old = 3, 89 islets scored; n3-month-old ctrl = 3, 91 islets scored; n3.5-month-old = 3, 93 islets scored; n18.5-month-old = 3, 83 islets scored; n19-month-old ctrl = 3, 83 islets scored; n19.5-month-old = 3, 88 islets scored; DT (2-week-old): n0.5 mpa = 6, 333 islets scored; n1 mpa = 3, 91 islets scored; n1.5 mpa = 3, 90 islets scored; DT (2-month-old): n0.5 mpa = 3, 76 islets scored; n1 mpa = 3, 77 islets scored; n1.5 mpa = 3, 81 islets scored; DT (1.5-year-old): n0.5 mpa = 3, 74 islets scored; n1 mpa = 3, 81 islets scored; n1.5 mpa = 3, 77 islets scored. Error bars show s.d. Welch’s test, one-way ANOVA (P < 0.001), Mann–Whitney (P = 0.0238). i, Ki67+ cells are hormone, chromogranin-A-negative; lineage-traced α- and DT-spared β-cells are Ki67-negative. Scale bars, 20 µm.
