Extended Data Figure 6. tyra-3 acts in the intestine to inhibit stress resistance.

a, Top, Gene structure of tyra-3. Coding sequences are represented by black boxes. Bottom, Confocal images of transgenic animals expressing mCherry driven by a short fragment of tyra-3 promoter (short promoter, 1.8 kb upstream of start codon). mCherry expression is limited to a subset of head and tail neurons (and vulval cells). Scale bar: 200 μM. A mCherry reporter driven by a long 3.4 kb promoter (Ptyra-3_long) show expression in both, neurons and intestine (see manuscript Fig 3b). b, Survival percentages of tyra-3 mutant animals expressing tyra-3 cDNA driven by Ptyra-3_long (Endogenous expression), Ptyra-3_short (Expression in a subset of neurons), Prgef-1 (Pan-neuronal) or Pelt-2 (Intestinal) promoter upon exposure to heat stress with or without tyramine (10 mM). Bars represent mean ± SEM, n=5, 80–100 animals/condition. For conditions without TA, One-way ANOVA followed by Holm-Sidak post-hoc test versus wild-type was used. For conditions with TA, two-tailed Student’s t test (versus same strain without TA) was used. Expression of tyra-3 in the intestine, but not in neurons, was sufficient to restore the stress sensitivity and the negative impact of exogenous tyramine on heat. c, Representative Kaplan-Meier survival curves of the wild-type, tyra-3 null mutants and animals expressing tyra-3 solely in the intestine(Pelt-2::tyra-3). Animals were food deprived as L4s in the absence or presence of 10 mM TA. The curves are representative of three independent replicates with similar results (n=3). 40–80 animals were used per condition per experiment. Two-tailed log-rank test was used for statistical comparison. Expression of tyra-3 in the intestine restores the negative impact of exogenous tyramine lifespan upon starvation d, SI to heat exposure (4 h, 35°C) of animals pre-exposed to vibrational stimulus (tapping). Intestinal expression of tyra-3 restores the detrimental effect of tapping on the stress response. Bars represent mean ± SEM, n = 3, 30–40 animals/condition. One-way ANOVA followed by Holm-Sidak post-hoc test for multiple comparisons.