Extended Data Fig. 9: One-carbon metabolism activation across 5 different cancer cachexia mouse models.
(a-g) Healthy controls (wildtype, Ctrl, grey) and cachectic APCMin mutant mice (intestinal cancer, pink). Mice were fasted for 6 h before tissue sampling. n = 5 animals per group. (h-n) Healthy controls (PBS-injected, Ctrl, grey) and cachectic LLC tumour-bearing mice (lung carcinoma, green). Tissue sampling in random-fed animals. n = 5 animals per group. (o-u) Healthy controls (wildtype, Ctrl, grey) and cachectic KPP mutant mice (pancreatic ductal adenocarcinoma, violet). Tissue sampling in random-fed animals. n = 4 animals per group. (a-c, h-j, o-q) Body weight loss expressed as percentage of initial or maximal body weight (a, h, o); final tissue weights: GC muscle(s) (b, i, p) and eWAT (c, j, q). (d-e, k-l, r-s) Relative mRNA expression level of key enzymes of one-carbon metabolism and related pathways in liver (d, k, r) and GC muscle (e, l, s). (f-g, m-n, t-u) Metabolite levels of one-carbon metabolism and related pathways in liver (f, m, t) and GC muscle (g, n, u). Metabolites are similar to the list presented in Fig. 3b. (v-ab) See also Fig. 7a for experimental setup. Mice were divided into 4 groups: mice injected with PBS, healthy controls, no tumour (Ctrl, grey, n = 6 animals per group); mice orthotopically injected with Panc02 pancreatic cancer cells, mild cachexia (Panc02-Mild, blue, n = 6 animals per group); mice orthotopically injected with 8025 pancreatic cancer cells divided into two groups, mild cachexia (8025-Mild, light green, n = 7 animals per group) and cachexia (8025-Cax, dark green, n = 5 animals per group). (v-w) Initial (v) and final (w) body, lean and fat mass. (x) Loss of body, lean and fat mass expressed as percentage of initial mass. (y) Final tissue weights (GC muscle, eWAT). (z) Final tumour weights. (aa) Relative mRNA expression of atrogens in GC muscle. (ab) Relative mRNA expression of key enzymes of one-carbon metabolism and related pathways in eWAT. Data expressed as fold change of Ctrl. Statistical analysis on raw data (2−ΔCt values and MS signal intensities, arbitrary units A.U.). Data are mean ± s.e.m. Statistical analysis: two-tailed, non-adjusted, Student’s t test or Mann and Whitney’s test (a-u), unpaired one-way ANOVA with Dunnett’s post-hoc tests or Kruskal Wallis with Dunn’s post-hoc tests vs. Ctrl (v-ab). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 compared to Ctrl animals.
