Figure 1. Expression of DAF-16::GFP. (a) Schematic of the
daf-16 gene adapted from [9, 10]. Solid lines represent genomic DNA, dashed lines represent spliced introns, white boxes represent 5' and 3' untranslated regions, blue boxes indicate fork head domain coding exons, and green boxes represent exons encoding green fluorescent protein (GFP). (b) DIC image of late embryo. (c) GFP fluorescence. DAF-16::GFP is first evident in many cells just before hatching. (d) GFP fluorescence of a fourth larval stage (L4) animal; anterior isleft. DAF-16::GFP is widely expressed, except in the pharynx (arrow) and germline. (e) DAF-16::GFP expression in a 20-day-old animal; anterior is left. Fluorescence is maintained in old animals and does not become localized under normal culture conditions either on agar plates or in liquid culture. (f) Size of IsDAF-16::GFP worms and controls; IsDAF-16::GFP animals are smaller at all stages. Small size is reversed by inhibition of
daf-16. (g) Daily fertility of IsDAF-16::GFP animals and controls. IsDAF-16::GFP animals have delayed and reduced fertility. Inhibition of
daf-16 results in more progeny earlier, even in the control group. Total average progeny produced +/- SEM: ExpRF4 on empty vector bRNAi = 278.6 +/- 8.3, ExpRF4 on
daf-16 bRNAi = 308.8 +/- 11.5, IsDAF-16::GFP on empty vector bRNAi = 169.5 +/- 23.1, IsDAF-16::GFP on
daf-16 bRNAi = 285.7 +/- 10.1. (f,g) Squares represent ExpRF4. Triangles represent IsDAF-16::GFP. Unfilled symbols and dashed lines indicate exposure to bacterial
daf-16 RNAi. Filled symbols and solid lines represent exposure to empty vector control bacterial RNAi. Each point is the average of ten animals, with standard error.