Extended Data Fig. 5: Cysteine-depletion mobilizes lipids for thermogenic response independently of thermoneutrality.
From: Cysteine depletion triggers adipose tissue thermogenesis and weight loss
a) Quantification of ATGL immunoblot shown in Fig. 3a, Actin was used as a loading control. b, c) Tissue lipidomics of brown adipose depot (BAT) from Cth−/− mice fed CTRL (n = 4) or CysF diet (n = 5) for 6 days with b) triglycerides (TG) and c) diacylglycerol species highlighted. AU: arbitrary units. d) Core body temperature (CBT) measured in the peritoneal cavity by implantation of Star-Oddi logger of Cth−/− mice fed with CTRL or CysF diet over 6 days and e) average day and night CBT of Cth−/− mice fed with CTRL or CysF diet. Recordings were taken every 30 min (n = 11 Cth−/− CTRL, n = 12 Cth−/− CysF, 3 independent experiments pooled). f) Fgf21 gene expression in the liver of Cth+/+ and Cth−/− mice fed CTRL or CysF diet for 6 days (n = 8 Cth+/+ CTRL, n = 10 Cth+/+ CysF, n = 8 Cth−/− CTRL, n = 12 Cth−/− CysF). g, h) Serum levels of g) FGF21 and h) GDF15 in Cth−/− and Cth−/−CHOP−/− mice after 5 days of CysF feeding, measured by ELISA (n = 9 Cth−/− and n = 7 Cth−/−CHOP−/−). i) SFAT, VFAT and BAT weight normalized to body weight of Cth−/− and Fgf21−/−Cth−/− mice after CysF feeding (n = 5/group). j) Respiratory exchange ratio (RER) of Cth−/− and Fgf21−/−Cth−/− mice upon CysF feeding, measured at day 3 and 4 in metabolic cages (n = 5/group). k) Ucp1 gene expression in SFAT of Cth−/− and Fgf21−/−Cth−/− mice after 6 days of CysF feeding (n = 11 Cth−/− and n = 12 Fgf21−/−Cth−/−). l) Representative H&E histology images of SFAT showing increased browning at day 6 in Cth+/+ and Cth−/− mice fed CysF diet and housed at 20 °C or at 30 °C. m) Ucp1 gene expression measured by qPCR in BAT of Cth+/+ and Cth−/− mice fed CysF diet and housed at 20 °C or at 30 °C for 6 days (n = 3 Cth+/+20 C, n = 4 Cth+/+30 C and Cth−/−20C, n = 5 Cth−/−30C). Data are expressed as mean±SEM. Box plots represent median value and extend to the 25th and 75th percentiles. Whiskers are plotted down to the minimum and up to the maximum value. Statistical differences were calculated by 2-way ANOVA with Sidak’s correction for multiple comparisons, and by unpaired two-tailed t-tests.
