(A–C) In vitro kinase activity of post-gravid or sterile adult worms, 6 days after the L4/adult molt. Kinase activity of cleared, sonicated lysates was assessed as γ-³²P-ATP incorporation per 20-µg protein sample, in 1 min at 30°C. Samples, quenched on ice, were electrophoresed on 10% polyacrylamide-SDS gels (Invitrogen). (A) Gels stained with SYPRO Ruby (Invitrogen) to confirm constant protein loads. (B) Image of ³²P β-emissions (Molecular Dynamics Storm) from the gel in A, dried under vacuum. (C) Data summary from 2–3 independent expansions each of N2DRM, age-1(hx546), age-1(mg44) F1 homozygotes (labeled age-1 mg44^F1), age-1(mg44) F2 homozygotes (labeled age-1 mg44^F2), and daf-16(m26) double mutants with each age-1 allele. These quantitations covered the full length of each lane, not all of which is shown in A and B. (D–F) Phosphoproteins, resolved by polyacrylamide gel electrophoresis and visualized by Pro-Q Diamond staining. (D) Image of total protein stained with Coomassie blue. (E) Image of fluorescence after Pro-Q Diamond staining; note that only the two phosphoprotein markers were detected. (F) Summary of data from 3 independent expansions of each strain at adult day 6. Significance: (C, F), P values are shown for 2-tailed t-tests (unequal variance) comparing each age-1 strain to N2DRM (values directly over bars) or comparing two strains connected by brackets (values over brackets).