The Principles of GLP help ensure the quality and accuracy of data in chemical testing, and help prevent scientific fraud, as adopted by the European Union (EU). We assessed pooled data by use of a random-effects model. All survival data were plotted via the Kaplan-Meier method. For RNAi experiments, animals were grown to the L4 stage on standard RNAi plates seeded with E. coli HT115 carrying an empty vector control (pAD12) or the relevant RNAi clone before transferring to PA14-seeded SK plates and assaying for survival at 25 °C. Exposing C. elegans to this treatment for 24 h at 15 °C before transferring to PA14 SK plates prevented the eggs from hatching. To analyze PA14 sensitivity of C. elegans strains at various stages of life, temporal assays were conducted by picking eggs of wild-type and tcer-1 strains on OP50 plates, growing at 20 °C and then transferring them onto PA14 SK plates at L4, day 2, day 4, day 6 or day 9 of adulthood and monitoring for survival at 25 °C as mentioned above. Reproductively active animals were transferred to fresh OP50 plates every day till the relevant day of PA14 exposure. For imaging age-related change in TCER-1::GFP levels, the IT213 strain was grown at 20 °C on OP50 till day 1 or 4 of adulthood before dissection.

Fertile strains were transferred every other day to fresh plates until progeny production ceased. For lifespan assays of strains with the temperature sensitive glp-1 mutation, eggs were picked and maintained at 20 °C for 2−4 h, transferred to 25 °C to induce sterility and then returned to 20 °C on day 1 of adulthood (72 h later) for lifespan analysis. For performing lifespan assays of transgenic strains with extrachromosomal arrays, eggs were picked onto fresh OP50 plates, incubated at the appropriate temperature and 48 h later L4 animals were screened under a Leica M165FC microscope with a fluorescence attachment (Leica Microsystems, Wetzlar, Germany) for animals carrying the red coinjection marker labeling pharyngeal muscles. Using a variation of this paradigm, PA01 seeded onto standard NGM plates was also used for pathogen sensitivity assays. To rule out the impact of internal hatching on experimental outcomes, wild-type and tcer-1 L4 larval stage animals were treated with 100 μg per ml of FUDR on NGM plates with OP50.

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