Extended Data Fig. 2: CID661578 does not induce β- or ductal-cell proliferation.

a-c, Representative confocal images of Tg(ins:H2BGFP);Tg(ins:flag-NTR) larvae at 6 dpf after β-cell ablation (3-4 dpf) and incubation with EdU together with DMSO (a) or 5 µM CID661578 (b) during the regenerative period (4-6 dpf). β-cells that proliferated are marked by EdU and displayed as yellow overlap (arrowheads). The number of β-cells that proliferated was not altered by CID661578 (c). Scale bar, 10 µm. n = 13 (Control) and n = 15 (CID661578). Unpaired two-tailed Student’s t test was used to assess significance for (c) ns=not significant (P = 0.8340). Data are presented as mean values ±SEM. The experiment shown in (a-b) has been repeated two times with similar results. d-g, CID661578 treatment does not alter the number or the proliferation of notch-responsive ductal cells. Confocal images of DMSO- (d) or CID661578-treated (e) Tg(tp1:GFP);Tg(ins:flag-NTR) larvae at 6 dpf after β-cell ablation and incubation with EdU. White dashed lines outline the pancreata of the larvae. Scale bar, 10 µm. Quantification of the number of notch-responsive cells (f) and the number of proliferating, EdU⁺ notch-responsive ductal cells (g) per larva is shown. n = 11 (Control) and n = 12 (CID661578). Data are presented as mean values ±SEM. h, Glucose levels of Tg(ins:flag-NTR) larvae treated with DMSO, cercosporamide or CID661578 during the regenerative period (4-6 dpf). Four larvae were pooled for each replicate. n = 4 (all treatments – 3 dpf), n = 5 (all treatments – 4 dpf) n = 5 (all treatments – 5 dpf), n = 4 (Control and Cercosporamide – 6 dpf) and n = 4 (CID661578– 6 dpf). 2way ANOVA test followed by Tukey’s multiple comparisons test was used to assess significance for *P = 0.0430 (5 dpf Control vs Cercosporamide), ****P < 0.0001 (5 dpf Control vs CID661578), ****P < 0.0001 (6 dpf Control vs CID661578) and ****P < 0.0001 (6 dpf Control vs Cercosporamide). Data are presented as mean values ±SEM.

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