(A) Caloric restriction has been shown to extend lifespan in many species (Kenyon, 2010). Thus, we tested if the memo-1(gk345) mutants are calorically restricted. A genetic model for caloric restriction in the worm has an Eat phenotype, i.e., this worm has a pumping defect in its pharynx, preventing the animal from eating efficiently (Lakowski and Hekimi, 1998). Since Pmemo::GFP is expressed in the pharyngeal area (Figure 1A and Figure 1—figure supplement 1C and l), we analyzed memo-1(gk345) mutants for a pharyngeal pumping defect as a readout of feeding rate. memo-1(gk345) mutants did not have altered pharyngeal pumping compared to wild type (N2) at the L4 stage (N = 10 per strain, P value 0.8747 determined with t-test unpaired two-tailed). (B) Loss of memo-1 does not affect progeny production. Brood size for wild type (N2) (268 ± 15) and for memo-1(gk345) mutants (292 ± 11) was not significantly different. Data represented as mean + s.e.m., P value = 0.1953 determined with t-test, unpaired, two-tailed. (C) Genes involved in the heat shock response, in the germ stem cell-less-mediated longevity (glp-1), and in the unfolded protein response (UPR) are not induced in memo-1(gk345) mutants, as determined by qRT-PCR for the indicated transcripts. N > 200 L4 animals, two merged independent trials in duplicates. xbp-1u = unspliced, xbp-1s = spliced. (D–F) Loss of memo-1 does not affect insulin/IGF-1 signaling. (D) Schematic of insulin/IGF-1 receptor signaling to DAF-16/FOXO in C. elegans. Under normal conditions, insulin/IGF-1 receptor (daf-2) signaling retains the FOXO transcription factor DAF-16 in the cytoplasm. When insulin/IGF-1 signaling is reduced, DAF-16 translocates into the nucleus to initiate transcription of target genes (sod-3, btb-14, lipl-4, mlt-1, lea-1). (E) Downstream targets of DAF-16/Foxo transcription factor that are upregulated when insulin/IGF-1 receptor (daf-2) signaling is reduced (rIIS) are not altered in memo-1(gk345) mutant animals. (F) DAF-16::GFP translocates into the nucleus with daf-2 knockdown (not shown), but not with memo-1 knockdown (N > 60 per condition; zIs356 [DAF-16::GFP]). (G) memo-1(gk345) mutants have higher mRNA levels of oxidative stress response genes gcs-1 and gst-4 compared to wild type (N2), as determined by qRT-PCR. For each condition, two biological samples in duplicates of 200 L4 worms each were analyzed by qRT-PCR. All data are represented as mean ± s.e.m. P values of * <0.05 relative to wild type (N2) control, determined by one sample t-test, two-tailed, hypothetical mean of 1. (H) memo-1 mutants are not heat stress resistant. Day one wild type (N2) and memo-1(gk345) mutants were placed at 35°C and scored every hour for survival (N = 100 per strain). There is no significant difference between wild type (N2) and memo-1(gk345) for heat stress survival determined with log-rank (Mantel-Cox) method to calculate P value = 0.1594.