Extended Data Fig. 6: Characterization of the Usp20KI/KI mice and analysis the metabolism of L-Usp20−/− and Usp20KI/KI mice on HFHS.
From: Feeding induces cholesterol biosynthesis via the mTORC1–USP20–HMGCR axis
a, Strategy to generate Usp20KI/KI mice expressing USP20(S132A/S134A). b–f, Eight-week-old male Usp20KI/KI mice and their male WT littermates under chow diet were subjected to fasting and refeeding (n = 5 per group). TC (b), TG (c), free FA (d), insulin (e) and glucose (f) in the serum were measured. g, Relative amounts of mRNAs in the livers of WT and Usp20KI/KI mice subjected to fasting and refeeding treatments measured by qPCR. Each value represents mean of data from five mice. h, i, In vitro HMGCR deubiquitination assay performed as depicted in Fig. 1b. j–n, Eight-week-old male L-Usp20−/− mice and their male WT littermates were randomly grouped (n = 6 per group) and allowed ad libitum access to water and the HFHS diet for 23 weeks as in Fig. 3a. j, Representative image of the mice after 23-week of HFHS diet. k, Cumulative food intake (n = 6 per group). l, Measurement of fecal energy by bomb calorimetry (n = 5 per group). m, n, Respiratory exchange ratio (RER) (m) and movement (n) as determined by metabolic cages (n = 6 per group). o–u, Eight-week-old male Usp20KI/KI mice and their male WT littermates were randomly grouped (n = 5 per group) and allowed ad libitum access to water and the HFHS Diet for 14 weeks. o, Representative images after 14-week of HFHS diet. p, Body weight (n = 5 per group). q, r, Whole-body composition (n = 5 per group). s, Liver weight (n = 5 per group). t, GTT (n = 5 per group). u, ITT (n = 5 per group). v, w, Effect of chronic HFHS diet. Eight-week-old male L-Usp20−/− mice and their male WT littermates were randomly grouped and allowed ad libitum access to water and normal chow diet or HFHS diet for 8 weeks. v, Mice were fasted for 4 h and then sacrificed. w, Mice were subjected to fasting and refeeding treatment as in Fig. 1a. Liver samples were analysed by western blotting. CD: chow diet; HFHS: high-fat and high sucrose diet; F: fasted; R: refed. Each value represents mean ± SEM. Experiments in (h, i, v, w) were performed as indicated twice with similar results. Statistical significance was determined using unpaired two-tailed Student’s t-test (b–f, l, m, q–s); or two-way ANOVA (n, p, t, u).
