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. 2023 Feb 16;42(6):e112202. doi: 10.15252/embj.2022112202

Figure 1. Intestinal inflammation promotes lipolysis and autophagy in adipose tissues.

Figure 1

  1. Schematic of experimental design. Sex‐matched and age‐matched wild‐type mice were treated for 5 days with 1.5–2% DSS in drinking water, before switched to water for two more days. Mice were sacrificed on day 7 post‐DSS induction.
  2. Body weight development upon DSS treatment (n = 13/group).
  3. Tissue weights measured in mesenteric (mWAT) and collective visceral white adipose tissue (visWAT), consisting of gonadal (gWAT), retroperitoneal and omental white adipose tissue on day 7 after start of DSS regime (n = 8/group).
  4. Circulating serum levels of FFA during DSS‐induced colitis on day 7 (n = 15/group).
  5. Representative immunoblot for key lipolytic enzymes HSL and ATGL protein expression and quantification (n = 5–6/group).
  6. Immunoblot analysis of autophagic flux in mWAT (upper panel) and gWAT (lower panel) adipose tissue stimulated ex vivo with lysosomal inhibitors 100 nM Bafilomycin A1 and 20 mM NH4Cl for 4 h or DMSO (Vehicle) (n = 3–4/group).
  7. Representative transmission electron microscopy images from mesenteric adipose tissue 7 days post DSS‐induced colitis induction. Lower panel is showing magnification of selected area. White arrows show autophagosomal structures.
  8. Atg8 homologs expression was measured by qPCR in visceral adipocytes fraction (left panel) and stromal vascular fraction (right panel) during DSS‐induced colitis (n = 7–8/group).
  9. Representative immunoblot for LC3‐I/‐II protein expression and quantification of autophagic flux in gWAT via ex vivo lysosomal inhibition using 100 nM Bafilomycin A1 and 20 mM NH4Cl for 4 h or DMSO (Vehicle). Mice were initially treated with 500 μg anti‐TNFα antibody or isotype control, before administrating DSS in drinking water for 5 days. Mice were sacrificed on day 7 post‐DSS induction (n = 5–6/group).
  10. Representative immunoblot for LC3‐I/‐II and ACTIN protein expression and quantification of autophagic flux in creeping fat tissues (CrF) and adjacent mesenteric adipose tissues (Ad. MAT) of Crohn's disease patients (n = 3/group). Additionally, autophagic flux was determined in the mesenteric adipose tissue (MAT) of a colorectal cancer patient as control (dotted line).

Data are represented as mean ± s.e.m. (B) Two‐Way repeated measures ANOVA. (C–E, G) Unpaired Student's t‐test. (I) Two‐Way ANOVA. (J) Paired Student's t‐test. *P < 0.05, **P < 0.01, ***P < 0.001.

Source data are available online for this figure.