<i>daf-2</i> and <i>daf-16a</i> are required in ADL, but not in the intestine, to regulate <i>srh-234</i> expression.

<p>A) Percentage of mutant animals with defects in insulin signaling expressing <i>srh-234</i>p::<i>gfp</i> at wild-type levels. <i>daf-2(e1307)</i> is a temperature sensitive allele. In all experiments, animals were raised at 15°C (permissive temperature) and shifted to the 25°C (restrictive temperature) as L4 larvae. Animals (n>150) were examined at 150× magnification for each genotype. B, C) Relative expression of <i>srh-234</i>p::<i>gfp</i> in <i>daf-2</i> or <i>daf-16</i> mutants carrying compared to wild-type when fed or starved. For strains carrying <i>ADL::daf-2</i> cDNA, <i>pan-neural::daf-2</i> cDNA, <i>intestine::daf-2</i> cDNA, <i>ADL::daf-16a</i> cDNA and <i>intestine::daf-16a</i> cDNA extrachromosomal arrays (see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004707#s4" target="_blank">Material and Methods</a>), data shown is for at least two independent transgenic lines. Animals (n = 18–25) were examined at 400× magnification for each genotype. * indicates values that are different from that of wild-type animals at <i>P</i><0.001, # and ## indicates the values that are different between the genotypes compared by brackets at <i>P</i><0.001, and <i>P</i><0.05, respectively, using either a two-sample <i>t</i>-test or a χ² test of independence. Error bars denote the SEM or SEP.