Journal: PLOS Biology

Article Title: The neurohormone tyramine stimulates the secretion of an insulin-like peptide from the Caenorhabditis elegans intestine to modulate the systemic stress response

doi: 10.1371/journal.pbio.3002997

Figure Lengend Snippet: (A) Representative epifluorescence images (20×) of L4 animals expressing Pins-3::GFP under basal conditions (20 °C on Nematode Growth Media plates seeded with OP50), under oxidative stress (1 mM FeSO 4, 2 h) or heat stress (30 °C, 6 h) and in the presence of exogenous tyramine (15 mM). Scale bar 50 µm. (B) Corresponding quantification of fluorescence levels per worm. Scatter dot plot with relative expression of Pins-3::GFP normalized to basal conditions of each independent experiment. Line at the median. n = 50−200 animals per condition (distributed across 3–4 independent experiments). One-way ANOVA and Dunn’s post hoc test vs. basal were used. * p < 0.05, **** p < 0.0001. (C) Log 2 fold-changes in ins-3 transcript levels in animals exposed to oxidative stress (1 mM FeSO 4 , 2 h) or heat stress (30 °C, 6 h) and in the presence of exogenous Tyramine (15 mM). Negative and positive values indicate down- and up-regulation of ins-3 , respectively. Fold change was calculated as ΔCt basal conditions/ΔCt test conditions. Results are shown as mean ± s.e.m. n = 4 independent experiments. The data underlying this figure can be found at https://osf.io/wfgvs/ .

Article Snippet: For ins-3 and ins-4 expression levels analysis, animals containing the corresponding transcriptional GFP reporter ( Pins-3::GFP and Pins-4:GFP , respectively) were imaged using an epifluorescence microscope (Nikon Eclipse TE2000-5) coupled to a CCD camera (Nikon DS-Qi2) with 20× objective.

Techniques: Expressing, Fluorescence

Journal: PLOS Biology

Article Title: The neurohormone tyramine stimulates the secretion of an insulin-like peptide from the Caenorhabditis elegans intestine to modulate the systemic stress response

doi: 10.1371/journal.pbio.3002997

Figure Lengend Snippet: Representative epifluorescence images (40×) of young not gravid adults expressing Pges-1::INS-3::VENUS in wild-type and tyra-3 null mutant background, in the absence or presence of exogenous tyramine (15 mM). Scale bar 25 µm. Fluorescence is observed in the two posterior coelomocytes of worms (inset). Right. Corresponding quantification of fluorescence intensity on the coelomocytes (normalized to wild-type animals without tyramine exposure). Line at the median. n = 35–40 animals per condition distributed across three independent experiments. For conditions with tyramine, a two-tailed Student’s t test vs. the same strain without tyramine was used. * p < 0.05. The data underlying this figure can be found at https://osf.io/wfgvs/ .

Article Snippet: For ins-3 and ins-4 expression levels analysis, animals containing the corresponding transcriptional GFP reporter ( Pins-3::GFP and Pins-4:GFP , respectively) were imaged using an epifluorescence microscope (Nikon Eclipse TE2000-5) coupled to a CCD camera (Nikon DS-Qi2) with 20× objective.

Techniques: Expressing, Mutagenesis, Fluorescence, Two Tailed Test