Fig. 4: Global cysteine deficiency induced adipose browning is UCP1 independent.
From: Cysteine depletion triggers adipose tissue thermogenesis and weight loss
a, Immunoblot analyses of CTH in the liver of male and female Cthf/f Alb:Cre− or Alb:Cre+ mice. b, Western blot detection of CTH in the SFAT of male and female Cthf/f Adipoq:Cre− or Adipoq:Cre+ mice. c,d, Serum cysteine and cystine determined by LC–MS/MS in Alb:CreCthf/f mice (n = 5 per group) (c) and Adipoq:Cre;Cthf/f mice (n = 4 CTRL and n = 5 CysF) (d) after 6 days of CTRL or CysF diet. AU, arbitrary units. e,f, Percentage body weight changes of Alb-Cre;Cthf/f mice (n = 5 Cthf/f CTRL, n = 6 Cthf/f CysF and n = 3 Alb-Cre;Cthf/f CTRL and CysF) (e) and Adipoq-Cre;Cthf/f mice (n = 5 per group) after 6 days of CTRL or CysF diet (f). g,h, Volcano plot of serum metabolites identified by LC–MS/MS in Alb-Cre;Cthf/f mice (n = 5 per group) (g) and Adipoq-Cre;Cthf/f mice (n = 4 CTRL and n = 5 CysF) (h) after 6 days of CTRL or CysF diet. Trans-sulfuration pathway related metabolites are highlighted in red. Significantly increased or decreased metabolites (−log10(P) >1.3 and ∣log2(FC)∣>1) are highlighted in blue and listed on the right. Cys, cysteine; Homocys, homocysteine; Met, methionine; SAH, S-adenosyl homocysteine. Supplementary Tables 3 and 6 provide the full list of metabolites. i–l, Cth−/− and Cth−/− Ucp1−/− mice were fed a CysF diet for 6 days (n = 8 per group). Per cent body weight change over 6 days of diet (i). Representative H&E histology images of SFAT after 6 days of diet (j). Energy expenditure measured in metabolic cages on days 4 and 5 of CysF diet (k). Linear regression analysis of EE against body mass during dark and light cycles at 4 and 5 days of weight loss (when adjusted to body mass covariate, EE of Cth−/− Ucp1−/− is significantly decreased, during both night and day) (l). m, CBTs measured in the peritoneal cavity by implantation of Star-Oddi loggers over 6 days of diet in male Cth−/− and Cth−/−Ucp1−/− mice fed CysF diet. Recordings were taken every 30 min and representative day 4 is plotted (n = 7 Cth−/−, n = 5 Cth−/−Ucp1−/−). n,o, Immunoblot staining of ATGL, TH and UCP1 in BAT of Cth−/− and Cth−/−Ucp1−/− fed a CysF diet for 6 days (n) and quantification using tubulin as loading control (o). p, Thermogenic markers gene expression analysis in BAT of Cth−/− and Cth−/−Ucp1−/− mice fed a CysF diet for 6 days, measured by qPCR (n = 8 Cth−/−, n = 10 Cth−/−Ucp1−/−). q, Gene expression of genes involved in futile creatine cycle in BAT of Cth−/− and Cth−/−Ucp1−/− mice fed a CysF diet for 6 days (n = 16 Cth−/−, n = 15 Cth−/−Ucp1−/−), quantified by qPCR. Data are expressed as mean ± s.e.m. Statistical differences were calculated by two-way ANOVA with Sidak’s correction for multiple comparisons or by unpaired two-tailed t-tests.
