A region between −500 bp and −405 bp in the lag-2 promoter is crucial for the DTC response to DAF-7/TGFβ. Expression levels of lag-2 low-copy GFP-PH reporters driven by (A-B) 2 kb, (C-D) 1 kb, (E-F) 500 bp and (G-H) 405 bp upstream in the DTC of wild-type, daf-7(e1372) and daf-7(e1372); daf-3(e1376) animals. Transgene alleles are naIs81, naIs84, naIs87 and naIs98, respectively. (B,D,F,H) Mean pixel intensity (arbitrary units), measured in the DTC as described in the Materials and Methods. The boxes indicate the minimum-maximum range of samples quantified. Scale bars: 5 μm. n.s. indicates P>0.05, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, two-tailed Student's t-test. n≥15 animals, one DTC scored per animal. Error bars represent s.e.m. In all cases, P>0.05 for wild type versus double mutants.