Figure 2. Plasma insulin level after feeding is dependent on time of day, not fasting duration.

(A) Experimental design of MSIS test. (B-C) Blood glucose level of male mice fasted for varying lengths of time, prior to and subsequent to feeding; FD axis indicates length of time since fasting initiation; red axis label indicates mice refed for 2 hours (B), and percent change in blood glucose level from fasted to fed state (C). (D-E) Plasma insulin level of male mice fasted for varying lengths of time, prior to and subsequent to feeding (D), and data presented with CR data shifted by 12 hours (E). (F) Experimental design of acute diazoxide treatment experiment. (G) Blood glucose level of male mice treated with either vehicle or diazoxide after a 12 hr fast (H-I) ATTs of ad libitum and CR-fed male mice (H) and AUC (I) after fasts of the indicated length. (B-E, G-I) n = 6–8 mice per diet per time point. (B-D) *p<0.05 AL-fed vs. CR-fed mice at each time point, (G) *p<0.05 AL-D vs. AL-V, #p<0.05 CR-D vs. CR-V, and $p<0.05 CR-D versus AL-D; Sidak’s test post two-way repeated measures ANOVA. (I) The overall effect of diet, fasting duration (FD), and the interaction represent the p-value from a two-way ANOVA. Data represented as mean ± SEM. AUC, area under the curve. See also Supplementary Figures 5.