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. 2021 Apr 23;19(4):e3001204. doi: 10.1371/journal.pbio.3001204

Fig 9. DAF-16/FoxO activity is distributed across multiple pharyngeal neurons to silence pharyngeal pumping in the dauer stage.

Fig 9

(A) DAF-16/FoxO depletion in pharyngeal neurons of daf-2(e1370) dauers using ehs-1prom4-driven TIR1. In 20/20 animals observed, AID-tagged DAF-16 was not detectable in TIR1-expressing neurons (all pharyngeal neurons except I1 and I4) after auxin treatment. Scale bars, 20 μm. (B) Pharyngeal pumping rate after DAF-16/FoxO depletion in daf-2(e1370) dauers using ehs-1prom4-driven TIR1 (otEx7628 and otEx7629). Horizontal lines represent median for 36 animals per condition. *** indicates p < 0.001 in Mann–Whitney test. (C) DAF-16/FoxO depletion in glutamatergic pharyngeal neurons of daf-2(e1370) dauers using eat-4prom7-driven TIR1. In 20/20 animals observed, AID-tagged DAF-16 was not detectable in TIR1-expressing pharyngeal neurons (I5 and M3) after auxin treatment. Scale bars, 20 μm. (D) Pharyngeal pumping rate after DAF-16/FoxO depletion in daf-2(e1370) dauers using eat-4prom7-driven TIR1 (otEx7670 and otEx7671). Horizontal lines represent median for 36 animals per condition. **** indicates p < 0.0001 in Mann–Whitney test. (E) DAF-16/FoxO depletion in cholinergic pharyngeal neurons of daf-2(e1370) dauers using unc-17prom4-driven TIR1. In 20/20 animals, AID-tagged DAF-16 was not detectable in TIR1-expressing pharyngeal neurons (I1, M1, M2, M4, M5, and MC) after auxin treatment. Scale bars, 20 μm. (F) Pharyngeal pumping rate after DAF-16/FoxO depletion in daf-2(e1370) dauers using unc-17prom4-driven TIR1 (otEx7668 and otEx7669). Horizontal lines represent median for 36 animals per condition. ** indicates p < 0.01 in Mann–Whitney test. (G) Neurotransmitter identity of all 14 pharyngeal neurons. Shaded boxes for each promoter fragment indicate the neurons in which TIR1 expression is strong enough to completely deplete AID-tagged DAF-16/FoxO in the presence of auxin (20 animals analyzed per strain). (H and I) Pharyngeal pumping rate in daf-2(e1370) dauers transferred from ethanol (solvent) to auxin or from auxin to ethanol after dauer entry (dauers were transferred on day 3 after hatching, and pumping rate was measured on day 6 after hatching). DAF-16/FoxO was depleted in the intestine (Pges1::TIR1) or in no tissue (no TIR1 control) in the presence of auxin. Horizontal red lines represent median for 18 animals per condition. p-Values are for Dunn’s multiple comparisons test performed after 1-way ANOVA on ranks. The data underlying this figure can be found in S1 Data. AID, auxin-inducible degron.