Figure S5. Radiometric assays indicated multiple kinases are commonly inhibited by the circadian period–lengthening compounds.

(A) “HotSpot” is a radiometric assay based on conventional filter-binding assays that directly measures kinase catalytic activity toward a specific substrate. ³³P ATP was used to measure ATP-competitive kinase inhibitions. (B) Activity profiles of different kinases in the presence of circadian period–lengthening compounds (100 μM). The DMSO (vehicle only) reactions were used as a full activity control, whereas the EDTA-only reactions are used as the no or zero activity control. Data are represented as mean ± SEM (n = 3). One-way ANOVA, Dunnett’s test. *** indicates P < 0.0001, ** indicates 0.0001 < P < 0.001, * indicates 0.001 < P < 0.01, and NS indicates P > 0.05.