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. 2019 Mar 8;15(3):e1007706. doi: 10.1371/journal.pgen.1007706

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© 2019 Harris et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 4 — (A-D) Mutating daf-1 that encodes the type I TGF-β receptor delays the decision to leave the OP50 lawn paired with 100% 2-nonanone (A, n = 8 and 9 assays for wild type and daf-1 mutant, respectively), and expressing the genomic DNA of daf-1 (B, n = 6 assays each) or a wild-type daf-1 cDNA in the RIM and RIC neurons (C, n = 6, 6 and 5 assays for wild type, transgenic animals and non-transgenic siblings, respectively) in the daf-1(m40) mutant animals rescues the delayed decision, but expressing wild-type daf-1 in the sensory neurons (D, n = 3, 5 and 2 assays for wild type, transgenic animals and non-transgenic siblings, respectively) does not rescue. Mutants are compared with wild type and transgenic animals are compared with non-transgenic siblings with Student t-test. (E-F) Inhibiting the activity of the RIM and RIC neurons by selectively expressing a histamine-gated chloride channel (E, n = 2 and 4 assays for wild type and transgenic animals, respectively) or blocking the synaptic release from these neurons by selectively expressing the tetanus toxin (F, n = 2 assays each) does not alter the decision to leave the OP50 food lawn paired with 100% 2-nonanone. Transgenic animals are compared with wild type. (G-I) Removing octopamine signaling in the daf-7(e1372) or daf-1(m40) mutants with a mutation that disrupts biosynthesis of octopamine tbh-1(ok1196) does not suppress the delayed leaving from the OP50 lawn paired with 100% 2-nonanone (G, n = 6 assays for wild type; n = 4 assays for daf-7 mutants; n = 2 assays for daf-1 mutants; n = 3 assays for daf-7;tbh-1 double mutants; n = 2 assays for daf-1;tbh-1 double mutants), but removing the tyramine and the octopamine signals with the mutation in tdc-1(ok914) in either the daf-7(e1372) (H, n = 6, 5 and 4 assays for wild type, daf-7 mutants and daf-7;tdc-1 double mutants, respectively) or the daf-1(m40) (I, n = 5, 4 and 5 assays for wild type, daf-1 mutants and daf-1;tdc-1 double mutants, respectively) mutant animals suppresses the delayed-decision phenotype in either of the mutant animals. Double mutants were compared with the respective single mutants using student’s t test. Each bar graph reports the average percentage of worms outside the lawn 15 minutes after the start of the assay. Mean ± SEM, ** p ≤ 0.01, *** p ≤ 0.001, n.s.; not significant.