Extended Data Fig. 1: In vivo chemical screen identifies stimulators of β-cell proliferation in zebrafish (related to Fig. 1).
From: In vivo screen identifies a SIK inhibitor that induces β cell proliferation through a transient UPR
(a) Validation of the LUCCI assay using NECA as a positive control in the basal state and during β-cell regeneration. Data in the basal state: N = 28 (DMSO), N = 13 (NECA 50 µM), and N = 32 (NECA 100 µM) wells. Data during regeneration: N = 30 (DMSO), N = 33 (NECA 50 µM), and N = 34 (NECA 100 µM) wells. One-way ANOVA with Tukey’s multiple comparisons test, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. Data are represented as the mean ± SEM. (b) Scatterplot of the efficacy of the hits identified during the basal state. PI3K: phosphoinositide 3-kinase. RTK: receptor tyrosine kinase. S/TK: serine/threonine kinase. (c) Dose-response activity of Lini and of HG. Tg(ins:NLuc-gmnn) or Tg(ins:NLuc-gmnn);Tg(ins:flag-NTR) larvae were treated with Lini or HG for 2 days (from 4 to 6 dpf), respectively. For Lini: N = 8 wells for X = −6.5; N = 11 wells for X = −6 and X = −5.3; N = 12 wells for X = −5; N = 15 for X = −5.7 and X = −4.7. For HG: N = 5 wells for X = −4.4, X = −4.7, X = −5 and X = −5.3. Data are represented as the mean ± SEM. A second order polynomial equation was used for modelling the bell-shaped curve. R2 = 0.13 for Lini and R2 = 0.32 for HG. (d) Effect of HG and Lini on the LUCCI signal under two conditions: in the basal state and during β-cell regeneration. Data in the basal state: N = 8 (DMSO), N = 8 (HG), and N = 6 (Lini) wells. Data during regeneration: N = 16 (DMSO), N = 10 (HG) and N = 5 (Lini) wells. One-way ANOVA with Tukey’s multiple comparisons test, ***P ≤ 0.001, ****P ≤ 0.0001. Data are represented as the mean ± SEM. (e) Number of β-cells in Tg(ins:H2B-GFP) larvae treated with DMSO or Lini from 4 to 6 dpf. Two-sided student’s t-test, ****P ≤ 0.0001. N = 23 (DMSO) and 9 (Lini) larvae. Data are represented as the mean ± SEM.
