Figure 2. Identification and validation of high-throughput screen lead compounds in murine islets at high and low glucose concentrations.

(A) Significant Z-scores (>3 standard deviations) and fold changes (>1.25-fold increase) for all 2640 screened compounds, with hit compounds indicated in blue. (B) Top 12 hit compounds identified from screen with a fold increase > 1.25 and a Z-score > 3, which were selected for further analysis. Known functions and published molecular pathways targeted by these compounds are indicated. (C) Model of potential mechanisms of action of the top 12 hit compounds to affect insulin secretion in the β-cell. (D) Glucose-responsive insulin secretion by ELISA at 2 mM and 20 mM glucose in WT mouse islets following exposure to four lead candidate compounds (n = 3–11 mice/compound). (E) Ivermectin (IVM) dose-response curve (n = 6–8 experimental repeats/dose), ranging from 0.078 µM to 80 µM IVM, in insulin-NanoLuciferase-expressing Beta-TC-6 cells. Shaded area represents 95% confidence intervals for the LOESS curve. All values represent mean ± SEM. **p<0.01, ***p<0.001.

Figure 2—figure supplement 1. High-throughput screen for modulators of insulin secretion in circadian mutant β-cells and validation of lead compounds.

(A) Compound exclusion flow chart delineating exclusion criteria and numbers of compounds excluded at each validation step. (B) Hit compound validation at concentrations of 10 and 100 µM in Bmal1^-/- insulin-NanoLuciferase cells (n = 3/compound). All values represent mean ± SEM. *p<0.05, **p<0.01, ***p<0.001.