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Mutant worms used in this study are unc-119 ed4 III unc-119 ed4 III)III.
Mutant worms used in this study are unc-119 ed4 III unc-119 ed4 IIII, and unc-45 e286 III unc-45 e286 III
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Whereas wild-type worms die and become rigid after many seconds, and take minutes to dissolve, we observed that cat-4 and ptps-1 mutant adult worms died in a few seconds or less, ruptured quickly, and their cuticles dissolved completely in <2 min. To quantify the cuticle fragility phenotype of cat-4 and ptps-1 mutants, we tested worms using a mild alkaline bleach treatment.
Forty 5-cm plates of lsy-12 mutants worms were used.
For smaller worms, use smaller hooks.
For thicker, longer worms use bigger hooks.
Besides the phenotype we have found in c30f12.4 mutant worms, we also determined, using IP and MS analysis, which C30F12.4 could interact with many proteins (Fig. S5 and Table S2).
For cDNA analyses, total RNA was isolated from N2 wild-type and mutant worms and reverse transcribed using a SuperScript III First-Strand Synthesis System kit (Invitrogen, catalog number: 18080-051).
Studies using null mutant worms revealed that both ACE-1 and ACE-2 are major functional enzymes with mutually compensating functions [11], [12] whereas ACE-3 does not compensate for the role of ACE-1 or ACE-2.
Murphy et al.[ 25] identified genes downstream of daf-16 in microarray experiments using adult daf-2; daf-16 hypomorphic mutant worms or worms in which DAF-2 and DAF-16 expression was knocked down with RNAi.
An unbiased approach to this question, using gene expression profiles from long-lived, IIS mutant worms, flies and mice, showed that cellular detoxification pathways were strongly up-regulated in all 3 organisms (McElwee et al., 2007).
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