Fig. 6: Characterization of brd-8.
From: A crucial role for dynamic expression of components encoding the negative arm of the circadian clock
a Race tube analysis of qa-2:brd-8V5 in the presence of QA at different concentrations as indicated. Strains were cultured on 0.1% glucose race tube medium on race tubes at 25 °C and synchronized by growth in constant light overnight (16-24 hrs) followed by transfer to darkness at the same temperature. b Plots of periods of qa-2:brd-8V5 in (a). Four or six biological replicates of each condition were run and error bars represent standard errors of the means. c Western blots showing BRD-8 levels in cultures of qa-2:brd-8V5 with different amounts of quinic acid grown in constant light at 25 °C. WT (328-4) and BRD-8V5 are the negative and positive controls respectively for the Western blots. d Western blotting of brd-8V5, wc-1V5, and esa-1V5 in cultures grown in different amounts of glucose (0.1 or 2%) in constant light. V5 was inserted to the C-termini of BRD-8, WC-1, and ESA-1 at their native loci respectively, and Western blotting against the same tag was used to fairly compare their protein levels. Red arrows point to BRD-8V5, WC-1V5, and ESA-1V5 as indicated. e Nuclear fractionation of BRD-8V5 from a culture grown in constant light. The nuclear fractionation was performed as previously described92; histone H3 and tubulin were followed as a nuclear and cytoplasm marker respectively. For c–e, similar results were observed from three replicates. f Bioluminescent analysis of luciferase expression under the control of promoters of brd-8, bye-1, eaf-3, vid-21, or histone h2a.z at 25 °C; LUC fused to the C terminus of BRD-8 ORF (translational fusion at the locus of brd-8) was also measured by the bioluminescent analysis at 25 °C. g Race tube analysis of brd-8-LUC with 0.1% glucose race tube medium in the dark at 25 °C. Source data were put in the Source Data file.
