Fig. 2: Cysteine depletion induces browning of adipose tissue.
From: Cysteine depletion triggers adipose tissue thermogenesis and weight loss
a, Representative images of subcutaneous (SFAT) and visceral (VFAT) fat sections stained for UCP1 from Cth−/− mice fed CTRL or CysF diet for 6 days (scale bar, 100 μm). b, Representative H&E-stained sections of SFAT of Cth−/− mice fed CTRL or CysF diet for 6 days or CysF diet followed by Cys-supplemented diet for 4 days (CysF + Cys) (scale bar, 100 μm). c, Western blot detection of ATGL and UCP1 in SFAT from Cth−/− mice after 6 days of CTRL or CysF diet or Cys supplementation after CysF-induced weight loss. Actin is used as a loading control. d, qPCR analysis of thermogenic genes in SFAT of Cth+/+ and Cth−/− mice fed CysF diet for 6 days (n = 8 Cth+/+ and n = 10 Cth−/−). e,f, Faecal calorie content (n = 6 per group) (e) and cumulative food intake of Cth−/− mice fed CTRL or CysF diet for 4 days (f) (n = 6 per group). g, Linear regression analysis of EE against body mass during dark cycle at 4 and 5 days of weight loss (n = 10 Cth+/+ CysF and n = 12 Cth−/− CysF). h, Per cent body weight change of Cth−/− mice fed with CTRL diet or CysF diet (red line) for 5 days and then switched to Cys-containing diet (orange line) for 3 days (n = 6 per group). i, RER measured in metabolic cages, of Cth−/− mice fed with CTRL diet (n = 6) or Cys-containing diet after CysF-induced weight loss (n = 4). j, Average food intake of Cth−/− mice fed with CysF diet and then switched to Cys-containing diet for 2 days (n = 7 per group). Significance was measured with paired t-test. k, Linear regression analysis of EE against body mass during dark cycle of Cth−/− mice fed with CTRL (n = 6) or Cys-supplemented diet after CysF-induced weight loss (n = 4), average values of the first two nights after diet switch. l, t-SNE plot of scRNA-seq showing cluster identities from SFAT SVF from Cth−/− mice fed CTRL or CysF diet at day 4 of weight loss and bar chart showing population fold change (FC) in relative abundance of each cluster comparing Cth−/− CysF versus Cth−/− CTRL. DC, dendritic cell; APC, antigen-presenting cell. m, t-SNE plot displaying Pdgfra expression in red across all populations and Monocle analysis of clusters 0, 1 and 2, with colouring by pseudotime to show right most cluster giving rise to two separate clusters. Each cluster represented by colour in Cth−/− CTRL and Cth−/− CysF. Data are expressed as mean ± s.e.m. Statistical differences were calculated by two-way ANOVA with Sidak’s correction for multiple comparisons or unpaired two-tailed t-tests. NS, not significant.
