Extended Data Fig. 1: Cysteine depletion induces weight-loss in mice without overt pathology.
From: Cysteine depletion triggers adipose tissue thermogenesis and weight loss
a) Cystathionine and homocysteine measurements by MS/MS in human SFAT at baseline (B) and after 12 months of caloric restriction (n = 14). AU: arbitrary units. b) Schematic of Cth−/− and Cthfl/fl mice generation (KOMP construct) used to cross to either Alb:cre or Adipoq:cre. c) Body weight of Cth+/+ and Cth−/− mice fed with CTRL or CysF diet for 6 days (n = 5 Cth+/+ CTRL, n = 6 Cth+/+ CysF, n = 4 Cth−/− CTRL, n = 5 Cth−/− CysF). d) Fat mass and lean mass measured by EchoMRI of male Cth+/+ and Cth−/− after 6 days of CTRL or CysF diet (n = 5 Cth+/+ CTRL, n = 12 Cth+/+ CysF, n = 8 Cth−/− CTRL, n = 17 Cth−/− CysF). e) Cth+/+ and Cth−/− mice were fed ad libitum (ad lib) or pair fed CTRL or CysF diet (n = 4 Cth+/+CysF ad lib, n = 5 Cth+/+CysF pair fed, n = 7 Cth−/−CTRL pair fed, n = 5 Cth−/− CysF pair fed). Percentage body weight change over 6 days of diet. f) Accumulated food intake of Cth+/+ and Cth−/− mice over 6 days of CysF feeding measured in metabolic cages (n = 10 Cth+/+ and n = 12 Cth−/−). Cage image and video show that Cth−/− mice on CysF diet at day 5 have normal activity. g) Qualitative assessment of nest building (score from 0 to 4) and presence (score=1) or absence (score=0) of kyphosis in WT and Cth−/− mice (n = 12/group). h) Gait assessment, ledge test and hindlimb clasping test were performed to measure motor coordination in WT and Cth−/− mice. Mice were scored from 0 (normal behaviour) to 1 (abnormal behaviour) (n = 12/group). i) Representative H&E-stained sections of kidney, lung, heart, and liver from female Cth−/− mice fed control diet or cystine-deficient diet for 6 days, lack notable pathologic changes and do not differ in microscopic changes by diet in the tissues examined. C = renal cortex, M = renal medulla A = airway, P = pulmonary artery, > = central vein, and * = portal triad. Kidney scale bars=200 μm, lung, heart, liver scale bars= 100μm. Data are expressed as mean±SEM. Statistical differences were calculated by 2-way ANOVA with Sidak’s correction for multiple comparisons, or by unpaired two-tailed t-tests. Panel b created with BioRender.com.
