Fig. 6. HLH-2 and LAG-1 bind to egl-43 regulatory regions. (A,B) EMSA with in vitro translated (TNT) LAG-1. Shifted complexes (B1 and B2) appear in the presence of LAG-1. The binding complexes are competed by DNAs containing the wild-type (w) but not the mutated (m) LAG-1 binding sites. `F' indicates the migration of the free DNA probe. (C,D) EMSA with purified HLH-2. A shifted complex `B' appears with HLH-2 and the wild-type ACEL-like probe, but not with Luciferase or with the probe on which both E-boxes are mutated. (D) The binding complex is competed by DNAs containing the wild-type (w) but not the mutated (m) E-boxes. DNA sequences are listed in Table 1; the numbers in B and D correspond to those of DNA competitors in Table 1. (E) PCR analysis from ChIPs performed with the extracts from animals expressing SEL-8::GFP. Pre-IP represents the input extracts subjected to IP. The 5'-region of lip-1, which contains four LAG-1 binding sites and was shown to be immunoprecipitated with LAG-3 (SEL-8) antibodies (Lee et al., 2006), was used as a positive control.