Figure 3. Transcriptional suppression of IIS genes in age-1(mg44) F2 adults.

Transcript levels were assayed by real-time polymerase chain reaction (RT-PCR). Expression histograms are shown superimposed on a simplified schematic of the IIS pathway. Yellow arrows indicate protein phosphorylations (symbolized by circled P's), and orange arrows indicate binding/activation by phosphatidylinositol 3,4,5-triphosphate (PIP₃, red “structural” symbols). Transcriptional outputs are indicated by open block arrows. Within each histogram, means±SEM are shown on a log(2) scale for steady-state transcript level, comparing wild-type to four age-1 mutant populations and to dauer larvae. For each strain indicated, fold changes are shown (e.g., “3×”), of age-1(mg44)-F2 relative to N2DRM. The age-1(mg44) worms are post-gravid F1 homozygotes at days 8–9 of adulthood, or F2 homozygotes at day 10 after the L4/adult molt; other strains (N2DRM, age-1(hx546), and daf-16(mu86); age-1(mg44) double mutants) were harvested as post-gravid adults (day 6 of adulthood), or as dauer larvae (N2DRM only, from starved, dense cultures 1 day after >98% of worms had become resistant to 1% sodium dodecyl sulfate). Transcript levels were assayed for 3–8 independent biological replicates, with two cDNA syntheses and two RT–PCRs for each. All results were normalized to the mean of three control genes (β-actin, T08G5.3, and Y71D11.3) that did not differ significantly among strains. Significant differences, relative to N2DRM controls, are indicated by asterisks in Table 1.