Fat utilization genes are direct targets of <i>C</i>. <i>elegans</i> DAF-12.

<p>(A, B) Gel electrophoresis mobility shift assays with [³²P]-labelled oligonucleotides corresponding to each of the candidate DAF-12 response elements (shown in the schematics of the promoters above the panels) for TG lipase (<i>K08B12</i>.<i>1</i>) and acyl-CoA synthase (<i>acs-3</i>). Gel shifts were performed in presence or absence of in vitro translated DAF-12 protein ± the presence of 20-fold and 200-fold (in A) or 200-fold (in B) excess unlabeled competitor oligonucleotides. ns, non-specific oligo at 200-fold excess. (C) Luciferase reporter assays with candidate response elements shown in (A,B) from HEK293 cells cotransfected with reporter and DAF-12 expression plasmids and treated with vehicle or 1 μM DA (n = 3 ± S.D.). (D) Nuclear run-on assays of <i>K08B12</i>.<i>1</i> and <i>acs-3</i> transcription in L2 larvae of <i>din-1;daf-9(dh127;dh6)</i> worms transiently treated by vehicle or 500 nM DA (n = 3 ± S.D.). *<i>P</i>< 0.05 comparing vehicle to DA treatment by Student’s t-test. Vehicle, ethanol; DA, Δ7-dafachronic acid.