Consistent with qRT-PCR results, the expression of GFP::
hlh-13 was regulated in response to DAF-16 in L1, L2/dauer, L4, and adult worms. L3 stage worms were not tested due to the dauer-constitutive phenotype of
daf-2(
e1370) at 25 C. Specifically, the GFP expression in the hypodermis at the head and the tail showed elevated expression in the
daf-2(
e1370) mutant compared to that in the
daf-16(mgDf47);daf- 2
(e1370) double mutant at 25 C. Also consistent with qRT-PCR, no difference in expression of the GFP::
hlh-13 transgene was observed in wild-type compared to the
daf-16(mgDf47) strain. In contrast to the hypodermal expression, the neuronal expression of
hlh-13 was not responsive to DAF-16 under conditions tested. These data together indicate that the neuronal expression of
hlh-13 is constitutive and is not under transcriptional control of IIS/DAF-16, whereas the hypodermal expression is regulated by IIS/DAF-16. This variation in transcriptional control may indicate multiple functions for
hlh-13, both dependent and independent of DAF-16.