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. 2010 Dec 14;8(12):e1000559. doi: 10.1371/journal.pbio.1000559

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Hirota et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) The chemical structure of compound 1 and longdaysin. (B) Effects of compound 1 and longdaysin on the luminescence rhythms in Bmal1-dLuc U2OS cells. Luminescence rhythms were monitored in the presence of various concentrations of compound (10 points of 3-fold dilution series in DMSO; final 0.7% DMSO). The representative luminescence profiles for longdaysin treatment are indicated as raster plot (left panel), in which each horizontal raster line represents a single well, with elapsed time plotted to the right. Luminescence intensity is indicated by color scale. Period parameter was obtained by curve fitting, and period change relative to the mean of DMSO control was plotted against compound concentration (right panel; the mean with SEM, n = 4). Longdaysin showed cytotoxicity at 71 µM. (C–E) Effect of longdaysin on the circadian period in adult tail fibroblasts (C), lung explants (D), and SCN explants (E) from mPer2^Luc knockin mice. Fibroblasts were cultured in the presence of various concentrations of longdaysin (the mean with SEM, n = 4). Lung and SCN explants were cultured in the presence of increasing concentration of longdaysin (0 to 9 µM; 1 wk for each concentration), and period change relative to the mean of DMSO control at first week was plotted for individual culture (n = 3 for lung and n = 2 for SCN).