Extended Data Figure 8. The flight response inhibits stress-dependent nuclear translocation of DAF-16.

a, Left, Fluorescence images of young adults expressing the translational reporter Pdaf-16::DAF-16a/b::GFP upon exposure to mild stressors as described in Fig. 1: 1 h 1 mM Fe²⁺ (Oxidation: Ox), 0.5 h 35°C (Heat: °C), 8 h food deprivation (Fasting: Fst), or tap stimulus every 5 min for 2.5 h (Tap). Fluorescence images of young adults expressing the translational reporter Pdaf-16::DAF-16a/b::GFP. Right, Scatter dot plot (line at the mean) with the number of cells with nuclear DAF-16 per animal. n for each condition is indicated in the figure. One-way ANOVA, Dunnett’s post-hoc test were used compared to naïve. Scale bar: 150 μm. b, Top, DAF-16 localization in wild-type and tdc-1 mutant animals exposed to vibrational stimulus (tap every 5 minutes) upon heat exposure (37°C, 1 h). Bottom, Scatter dot plot (line at the mean) with the number of cells with nuclear DAF-16 per animal (normalized to naïve animals (left). n for each condition is indicated in the figure. Two-tailed Student’s t-test was used versus same strain without tapping. Scale bar: 150 μm. Repetitive induction of the flight response impairs DAF-16 localization to the nucleus in wild-type, but not in tdc-1 mutant animals. c, Top, DAF-16 localization upon heat exposure (37°C, 1 h) of RIM::ChR2 transgenic animals raised with or without all-trans retinal (ATR) subjected to 5 second 617 nm light pulses every 5 minutes. Bottom, Scatter dot plot (line at the mean) with the number of cells with nuclear DAF-16 per animal (normalized to ATR-treated animals). n for each condition is indicated in the figure. Two-tailed Student’s t-test was used. Scale bar: 150 μm. Repetitive optogenetic activation of the tyraminergic neuron RIM impairs DAF-16 nuclear translocation.