File S1 - Fibroblast Growth Factor 21 Is Not Required for the Reductions in Circulating Insulin-Like Growth Factor-1 or Global Cell Proliferation Rates in Response to Moderate Calorie Restriction in Adult Mice

Contains the following files: Figure S1. Time course of plasma GH levels in CR and AL mice. Plasma GH levels in A) AL and B) CR mice over 5.25 h time course experiment (n = 8 per diet). Blood was collected from each mouse via the tail vein every 45 min. Each line represents the plasma GH levels over the time course experiment for a single mouse. CR mice were fed at their regular feeding time (1600 h). Figure S2. Time course of plasma GH levels in CR and AL mice. Plasma GH levels in AL and CR mice at A) 1530 h, B) 1615 h, C) 1700 h, D) 174 5h, E) 1830 h, F) 1915 h, G) 2000 h and H) 2045 h (n = 8 per diet). Blood was collected from each mouse via the tail vein every 45 min. CR mice were fed at their regular feeding time (1600h). Student's unpaired two-tailed t-tests were used for all between-group analyses. Figure S3. Blots used to quantify hepatic ph-total JAK2 and STAT5 levels in AL and CR mice. Hepatic protein levels of A–B) Total-JAK2, C–D) ph-JAK2, E–F) Total-STAT5 and G–H) ph-STAT5. * = One or more bands associated with ph/total quantification was not assessed or was imperfect. Blots were cut at indicated locations prior to primary antibody incubation. Figure S4. Ph-total hepatic STAT5 levels in AL-fed and CR-fed WT and FGF21-KO mice. n = 4–5 per genotype per diet per time point. Data normalized to WT/AL mice at 1500 h. All between-group analyses were performed using a two-way ANOVA with a Bonferroni post hoc test at each time point. Figure S5. Blots used to quantify hepatic ph-total STAT5 levels in AL-fed and CR-fed WT and FGF21-KO mice. Hepatic protein levels of A) Total-STAT5 in WT/AL and WT/CR mice at 1500 h, B) ph-STAT5 in WT/AL and WT/CR mice at 1500 h, C) Total-STAT5 in KO/AL and KO/CR mice at 1500 h, D) ph-STAT5 in KO/AL and KO/CR mice at 1500 h, E) Total-STAT5 in WT/AL and WT/CR mice at 1900 h, F) ph-STAT5 in WT/AL and WT/CR mice at 1900 h, G) Total-STAT5 in KO/AL and KO/CR mice at 1900 h and H) ph-STAT5 in KO/AL and KO/CR mice at 1900h. + con = extract from UT-7 cells treated with GM-CSF (positive control for STAT5 phosphorylation). Blots were cut at indicated locations prior to primary antibody incubation. (PDF)