Additional file 1 of ASGR1 deficiency diverts lipids toward adipose tissue but results in liver damage during obesity
Creators: name: Svecla, Monika
name: Da Dalt, Lorenzo
name: Moregola, Annalisa
name: Nour, Jasmine
name: Baragetti, Andrea
name: Uboldi, Patrizia
name: Donetti, Elena
name: Arnaboldi, Lorenzo
name: Beretta, Giangiacomo
name: Bonacina, Fabrizia
name: Norata, Giuseppe Danilo
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Medicine
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Mental Health
subject: Virology
subject: Computational Biology

description: Additional file 1: Figure S1. Weight and energy metabolism. A Total body weight after 20 weeks of high-fat diet (HFD). B Spleen and C Mesenteric lymph nodes (mLN) after 20 weeks of HFD. Each bar and error represent the mean ± SEM. Figure S2. Circulating glucose and lipid homeostasis. A Basal glucose after 14h of fasting before glucose tolerance test (GTT) in WT and ASGR1−/− mice after 20 weeks of high-fat diet (HFD). B Basal glucose after 4h of fasting before insulin tolerance test (ITT) in WT and ASGR1−/− mice after 20 weeks of HFD. C Triglycerides levels during the oral fat load and D after poloxamer injection in WT and ASGR1−/− mice after 20 weeks of HFD. E Plasma aspartate aminotransferase (AST) levels and F plasma alanine transaminase (ALT) in WT and ASGR1−/− mice after 20 weeks of HFD. Each bar and error represent the mean ± SEM. Figure S3. Visceral adipose tissue metabolism. A Principal component analysis (PCA) of the VAT proteome of WT and ASGR1−/− mice after 20 weeks of HFD. B Percentage of differently expressed proteins in VAT samples of WT and ASGR1−/− mice after 20 weeks of high-fat diet (HFD). C, D Gene expression of transcription factors and LXR-targets. E Inhibited metabolic pathways (z-score &lt; -2) and Factivated metabolic pathways (z-score &gt; 2) for the significantly differentially expressed proteins in WT and ASGR1−/− VAT after 20 weeks of HFD. G Lipogenesis index (C16:0/C18:2), (H) CE/FC ratio and (I) Phospholipid content (nmol/μg proteins) in WT and ASGR1−/− VAT after 20 weeks of HFD. Each bar and error represents the mean ± SEM. *p &lt; 0.05, **p &lt; 0.01 and ***p &lt; 0.001. Figure S4. Flow cytometry gating strategy for the analysis of innate immune cells. A Doublets were first excluded by the analysis (FSC-H vs. FSC-A). Leukocytes were gated based on dimension (FSC-H vs SSC-H) and CD45 positivity. Immune cells were identified as follows: B cells (CD19+, CD3−), T cells (CD19−,CD3+), macrophages (CD19−,CD3−,F4/80+), Natural killer cells (NK, CD19−,CD3−,F4/80−,NK1+), monocytes (NK1−,CD11b+,GR-1+ SSC-Hlow among F4/80− cells), neutrophils (NK1−,CD11b+,GR-1+,SSC-Hhigh among F4/80− cells), regulatory NK cells (CD27+,CD11b− among NK1+), cytotoxic NK cells (CD27+,CD11b− among NK1+), dendritic cells (DC, CD11c+ among F4/80− cells). Figure S5. Flow cytometry gating strategy for the analysis of adaptive immune cells. A Cells were gated on dimension (FSC-H vs SSC-H) and single cells discriminated (FSC-H vs. FSC-A). Leukocytes were then identified as CD45+ and T cells as CD3+ cells (CD45+, CD3+). Among CD45+CD3+ cells, CD4+ (T helper) and CD8+ (T cytotoxic) T cells were selected. Figure S6. VAT and liver immunophenotyping. The experiments are performed in three experimental groups (n = 6–14), after 20 weeks of high-fat diet (HFD). Immunophenotypic analysis through flow cytometry of VAT in WT and ASGR1−/− mice after 20 weeks of HFD: cells per gram of tissue of T helper cells (CD4+) A, cytotoxic T cells (CD8+) B and B cells (C). D–H Immunophenotypic analysis through flow cytometry of the liver in WT and ASGR1−/− mice after 20 weeks of HFD: D Monocytes E Macrophages F Neutrophils G CD 4+ T cells H CD 8+ T cells expressed as cells per gram of tissue. Each bar and error represent the mean ± SEM. Figure S7. Liver lipids and metabolism. (A) Liver cholesterol levels and (B) Liver triglyceride levels in WT and ASGR1−/− mice after 20 weeks of HFD. (C–E) Heatmap with Pearson clustering representing the oxidative phosphorylation pathway (FDR &lt; 0.01) subdivided in C Mitochondrial complex I (D) Mitochondrial complex II, III and IV (E) Mitochondrial complex V.