Luciferase reporter assays

Unless specified otherwise in the figure legends, all Neurospora strains were inoculated in 96-well plates and grown at 25° in 12 hr light, 12 hr darkness (LD) cycle for 72 hr before being transferred DD for luciferase analyses. The luciferase reporter assay analyses were performed as reported (Larrondo et al. 2015). Briefly, 96 well plates were inoculated with LN-CCD media (1× Vogel’s salts, 0.03% glucose, 0.05% arginine, 50 ng/ml biotin, 1.5% agar), supplemented with QA 0.01 M and 12.5 µM luciferin. Luciferase signals were acquired with a PIXIS 1024B camera (Princeton Instruments) and quantified with a customized macro developed for the ImageJ software; luminescent signals were accumulated for 5 min, three times per hour over the time course in DD. For metabolic compensation assays, glucose concentration was changed as indicate in the text.