Fig. 6

The contribution of NPAS3 domains to gene regulatory function. a, b Luciferase reporter assay demonstrating the gene-regulatory effects of NPAS3 domains. The C-terminus of NPAS3 is required to transactivate expression of a reporter construct, and without the bHLH and PAS domains the C-terminus drives non-specific transactivation. Reporter used is luciferase (luc2) expression driven by a construct including 797 bp proximal to the transcription start site of VGF. RLU = relative luciferase units, the ratio of firefly luciferase and renilla luciferase to normalize for transfection efficiency. Top and bottom of each box indicate the 25th and 75th centile, internal bar indicates the median, whiskers indicate data extremes. Data are representative of three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001. c Expression of NPAS3 domain constructs. Expected sizes: full length: 101 kDa, bHLH–PAS 49.4 kDa, PAS 35.8 kDa, PAS-TAD 98.2 kDa, TAD 53.2 kDa