To determine the specificity of Pitx2 activation of
unc-25/GAD,
unc-30(
e596) worms were co-injected with the Punc-30::Pitx2 expression con- struct along with either a wild-type Punc-25::GFP reporter or a mutant Punc-25::GFP reporter in which the UNC-30 binding sites were altered to CG-rich sequences. GFP expression was only detected from the wild type
unc-25 reporter, supporting that Pitx2 transactivation is dependent on intact UNC-30 binding sites in the
unc-25/GAD promoter.