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. Author manuscript; available in PMC: 2019 Sep 25.
Published in final edited form as: Nat Chem Biol. 2019 Mar 25;15(5):453–462. doi: 10.1038/s41589-019-0243-4

Figure 2. Phenotypic screening identifies SH-directed inhibitors that extend lifespan in C. elegans.

Figure 2.

(a) 95 SH-directed inhibitors were screened for their effect on C. elegans lifespan. Screening was done in 96-well plates and compounds were generally screened at 50 μM with a few exceptions that were screened from 5–25 μM due to lower solubility. Approximately 10 animals were incubated per well in liquid culture throughout the assay. Data for representative hit compounds (red dots) are shown as average values from multiple replicates (n = 3 independent experiments, shown on same vertical line, where black horizontal dash marks the average lifespan change for each hit compound). (b) The SH-directed inhibitor JZL184 (1) substantially extends lifespan. Shown is a representative Kaplan-Meier plot revealing ~45% lifespan extension by JZL184 (50 μM) (DMSO n = 80 worms, JZL184 n = 97 worms, P = 1.64E-10, compared to DMSO control). (c) Dose-dependent lifespan increase caused by JZL184. Half-maximal effect (EC50) of JZL184 was calculated to be 6 μM. Data represents mean ± s.e.m. (n = 11 independent experiments). (d) Representative lifespan curves demonstrating the interaction of JZL184 (50 μM) with known aging pathways: glp-1 (DMSO n = 57 worms, JZL184 n = 80 worms, P = 0.8359 compared to glp-1 DMSO control), hsf-1 (DMSO n = 74 worms, JZL184 n = 61 worms, P = 0.2471 compared to hsf-1 DMSO control), eat-2 (DMSO n = 75 worms, JZL184 n = 76 worms, P = 0.5207 compared to eat-2 DMSO control), xbp-1 (DMSO n = 73 worms, JZL184 n = 58 worms, P = 0.2121 compared to xbp-1 DMSO control), daf-16 (DMSO n = 81 worms, JZL184 n = 72 worms, P = 3.79E-11 compared to daf-16 DMSO control) and hif-1 (DMSO n = 79 worms, JZL184 n = 85 worms, P = 4.53E-13 compared to hif-1 DMSO control). For b and d, results are representative of 2 (glp-1) or 3 (all other conditions) independent experiments. Two-sided Mantel-Haenszel version of the log-rank test performed relative to DMSO control for b and d. Statistics for b-d are provided in Supplementary Dataset 2. (e) Food intake per worm in animals treated with DMSO or JZL184 (50 μM). Data represents mean ± s.e.m. (n = 144 wells per condition, P = 0.3752, two-sided unpaired t test, from 6 independent experiments).