Journal: Clinical and Molecular Hepatology

Article Title: Macrophage ATG16L1 expression suppresses metabolic dysfunction-associated steatohepatitis progression by promoting lipophagy

doi: 10.3350/cmh.2024.0107

Figure Lengend Snippet: Pharmacological enhancement of ATG16L1 expression prevents MASH development. The protein expression levels of STING signaling components in activated WT BMDMs treated with CM from PAOA-stimulated hepatocytes with or without the ATG16L1 enhancer peretinoin following DMXAA stimulation. (B) The protein levels of p-JNK, JNK, c-Jun, and c-Fos in activated WT BMDMs treated with CM. (C) The proinflammatory protein expression levels, (D) proinflammatory gene expression levels, and (E) Ifnb1 and (F) Tgfb1 mRNA expression in activated WT BMDMs treated with CM. (G) Schematic representation of HFHCD-fed or MCD-fed WT mice supplemented with dietary peretinoin. (H) Hepatic ATG16L1 protein expression levels. (I) Hepatic H&E staining, oil red O staining, Sirius red staining, and α-SMA immunohistochemistry. (J) NAS, serum ALT levels, and hepatic TG levels. (K) Hepatic Acta2, Col1a1, and Timp1 gene expression; n=6 mice/group. (L) Hepatic α-SMA, collagen- I, and TIMP-1 protein expression levels. (M) Hepatic Tnfa , Il6 , and Il1b gene expression levels. (N) Hepatic F4/80 + cell immunohistochemistry. (O) Representative eWAT H&E staining and F4/80 immunohistochemistry image of HFHCD-fed WT mice treated with or without peretinoin. MASH, metabolic dysfunction-associated steatohepatitis; ATG16L1, autophagy-related protein 16-like 1; BMDMs, bone marrow-derived macrophages; STING, stimulator of interferon genes; PAOA, palmitic acid- and oleic acid; CM, conditioned media; HFHCD, high-fat and high-cholesterol diet; MCD, methionine- and choline-deficient diet; NAS, NAFLD activity score; ALT, alanine aminotransferase; TG, hepatic triglyceride; eWAT, epididymal white adipose tissue. The data are expressed as the mean±SD. * P <0.05, ** P <0.01 (unpaired t test or ANOVA).

Article Snippet: Mice were randomly assigned to receive a normal chow diet (NCD), a high-fat and high-cholesterol diet (HFHCD) (Research Diets, Inc., New Brunswick, CA, USA) for 16 weeks (n=6–8 per group), or a methionine- and choline-deficient (MCD) diet (Research Diets, Inc.) for 6 weeks (n=6–8 per group).

Techniques: Expressing, Staining, Immunohistochemistry, Derivative Assay, Activity Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Critical and Diverse Roles of CD4 – CD8 – Double Negative T Cells in Nonalcoholic Fatty Liver Disease

doi: 10.1016/j.jcmgh.2022.02.019

Figure Lengend Snippet: Proportion of DNT cells increased in MCD-induced NAFLD for 5 weeks. ( A ) Percentages of DNT cells in CD3 + T cells among the liver, spleen, blood, ALNs, DLNs, MLNs, and ILNs from normal mice. ( B ) Representative H&E staining and Oil Red O and α-SMA staining in livers of NCD- and MCD-fed mice. ( C ) NAFLD activity score (NAS) in livers of NCD and MCD mice. ( D ) Serum ALT and serum AST were measured in NCD- and MCD-fed mice. ( E ) Representative flow cytometry plots and statistical analysis of percentages of intrahepatic DNT cells in CD3 + T cells and CD45 + T cells of NCD- and MCD-fed mice. ( F ) Numbers of intrahepatic DNT cells in NCD- and MCD-fed mice. ( G ) Representative flow cytometry plots and statistical analysis of annexin V + and Ki67 + expression in intrahepatic DNT cells in NCD- and MCD-fed mice. ( H ) Percentages of DNT cells in CD3 + T cells among the spleen, blood, ALNs, DLNs, and MLNs of NCD- and MCD-fed mice. n ≥ 4 mice/group. Two-sided P values <.05 were considered significant. ∗∗ P < .01; ∗ P < .05.

Article Snippet: After being allowed to adapt for 5 days, the C57BL/6 mice were fed either a NCD or a MCD (10401; Beijing HFK Bioscience, Beijing, China) for 5 weeks or a CDHFD for 18 weeks.

Techniques: Staining, Activity Assay, Flow Cytometry, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Critical and Diverse Roles of CD4 – CD8 – Double Negative T Cells in Nonalcoholic Fatty Liver Disease

doi: 10.1016/j.jcmgh.2022.02.019

Figure Lengend Snippet: RNA-sequencing showed the differentially expressed genes and biological function of DNT cells from livers of MCD-fed mice compared with NCD-fed mice. ( A ) Differences of genes with up-regulated (422) and down-regulated (458) expression in intrahepatic DNT cells from MCD-fed mice compared with NCD-fed mice; fold change ≤ 0.5 or fold change ≥ 1.5; P < .05. ( B and C ) Biological Process and KEGG pathway analyses were performed on the basis of the genes with significantly up-regulated and down-regulated expression in intrahepatic DNT cells from MCD-fed mice compared with NCD-fed mice. ( D ) GSEA of positive regulation of the T-cell apoptotic process in intrahepatic DNT cells from MCD-fed mice compared with NCD-fed mice. ( E and F ) Heatmap showing up-regulated and down-regulated genes related to cell apoptosis in intrahepatic DNT cells from MCD-fed mice compared with NCD-fed mice. The significantly changed genes related to cell apoptosis were confirmed via real-time PCR. ( G ) GSEA of cell killing in intrahepatic DNT cells of MCD-fed mice. ( H ) Heatmap showing the genes with down-regulated expression related to cell-mediated cytotoxicity in intrahepatic DNT cells from MCD-fed mice compared with NCD-fed mice. ( I ) The significantly changed genes related to cell-mediated cytotoxicity were confirmed via real-time PCR. ( J and K ) Representative flow cytometry plots and statistical analysis of GZMB and NKG2D expression in intrahepatic DNT cells from NCD- and MCD-fed mice. ( L ) GSEA of the inflammatory response in intrahepatic DNT cells from NCD- and MCD-fed mice. ( M ) Heatmap showing genes with up-regulated and down-regulated expression related to the inflammatory response in intrahepatic DNT cells from livers of MCD-fed mice compared with NCD-fed mice. ( N ) The significantly changed genes related to the inflammatory response were confirmed via real-time PCR. ( O ) GSEA of the response to IL17 in intrahepatic DNT cells from NCD- and MCD-fed mice. ( P ) Representative flow cytometry plots and statistical analysis of IL17A expression in intrahepatic DNT cells from NCD- and MCD-fed mice. n ≥ 5 mice/group. Two-sided P values <.05 were considered significant. ∗∗ P < .01; ∗ P < .05.

Article Snippet: After being allowed to adapt for 5 days, the C57BL/6 mice were fed either a NCD or a MCD (10401; Beijing HFK Bioscience, Beijing, China) for 5 weeks or a CDHFD for 18 weeks.

Techniques: RNA Sequencing, Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Critical and Diverse Roles of CD4 – CD8 – Double Negative T Cells in Nonalcoholic Fatty Liver Disease

doi: 10.1016/j.jcmgh.2022.02.019

Figure Lengend Snippet: Immunosuppressive function of DNT cells was controlled by TCRαβ+ DNT cells, and secretion of inflammatory factors was controlled by TCRγδ + DNT cells. ( A ) Representative flow cytometry gating strategy of TCR proportions in intrahepatic DNT cells. ( B ) Representative flow cytometry plots of TCR proportions in intrahepatic DNT cells from MCD- and NCD-fed mice. ( C ) Statistical analysis in apoptosis of TCR subtypes in intrahepatic DNT cells from MCD- compared with NCD-fed mice by flow cytometry. n = 3 mice/group. ( D ) Statistical analysis by flow cytometry of CXCR6 expression in liver and spleen TCRγδ + DNT cells and TCRαβ + DNT cells among NCD- and MCD-fed mice. ( E ) The mRNA expression of Cxcl16 in liver among NCD- and MCD-fed mice. ( F , H , and I ) Representative flow cytometry plots and mRNA of GZMB expression in intrahepatic TCRγδ + DNT cells and TCRαβ + DNT cells among NCD- and MCD-fed mice. ( G , J , and K ) Representative flow cytometry plots and mRNA of IL17A expression in intrahepatic TCRγδ + DNT cells and TCRαβ + DNT cells from NCD- and MCD-fed mice. n ≥ 5 mice/group. Two-sided P values <.05 were considered significant. ∗∗ P < .01; ∗ P < .05.

Article Snippet: After being allowed to adapt for 5 days, the C57BL/6 mice were fed either a NCD or a MCD (10401; Beijing HFK Bioscience, Beijing, China) for 5 weeks or a CDHFD for 18 weeks.

Techniques: Flow Cytometry, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Critical and Diverse Roles of CD4 – CD8 – Double Negative T Cells in Nonalcoholic Fatty Liver Disease

doi: 10.1016/j.jcmgh.2022.02.019

Figure Lengend Snippet: TCRγδ + DNT depletion resulted in lowered liver IL17A levels and significantly alleviated diet-induced NAFLD. ( A ) Representative flow cytometry plots of DNT proportions in TCRγδ + T cells. ( B ) Statistical analysis of TCRγδ + T cells in CD3 + T cells from the spleen and liver of NCD- and MCD-fed mice and TCRγδ + DNT-depleted mice. ( C and D ) Representative flow cytometry plots and statistical analysis of TCRγδ + T cells in intrahepatic DNT cells of NCD- and MCD-fed mice and TCRγδ + DNT-depleted mice. ( E ) Representative H&E staining and Oil Red O and α-SMA staining in livers of NCD- and MCD-fed mice and TCRγδ + DNT-depleted mice. ( F ) NAFLD activity score (NAS) in livers of NCD- and MCD-fed mice and TCRγδ + DNT-depleted mice. ( G and H ) Serum ALT and serum AST were measured in livers from NCD- and MCD-fed mice and TCRγδ + DNT-depleted mice. ( I–L ) Representative flow cytometry plots and statistical analysis of GZMB and IL17A expression in intrahepatic DNT cells from NCD- and MCD-fed mice and TCRγδ+ DNT-depleted mice. ( M ) Statistical analysis of interferon γ expression in intrahepatic DNT cells from NCD- and MCD-fed mice and TCRγδ + DNT-depleted mice by flow cytometry. ( N ) Proportions of monocytes (CD45 + Ly6G - F4/80 int CD11b hi ), Kupffer cells (CD45 + Ly6G - F4/80 hi CD11b int ), M1 monocytes (CD45 + Ly6G - F4/80 int CD11b hi CD11c + CD206 - ), and M2 monocytes (CD45 + Ly6G - F4/80 int CD11b hi CD11c - CD206 + ) among intrahepatic DNT cells from NCD- and MCD-fed mice and TCRγδ + DNT-depleted mice. n ≥ 5 mice/group. Two-sided P values <.05 were considered significant. ∗∗ P < .01; ∗ P < .05.

Article Snippet: After being allowed to adapt for 5 days, the C57BL/6 mice were fed either a NCD or a MCD (10401; Beijing HFK Bioscience, Beijing, China) for 5 weeks or a CDHFD for 18 weeks.

Techniques: Flow Cytometry, Staining, Activity Assay, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Critical and Diverse Roles of CD4 – CD8 – Double Negative T Cells in Nonalcoholic Fatty Liver Disease

doi: 10.1016/j.jcmgh.2022.02.019

Figure Lengend Snippet: Adoptive transfer of TCRαβ + DNT cells prevented the development and progression of NAFLD. ( A ) Serum ALT and serum AST levels were measured in NCD- and MCD-fed mice and TCRαβ + DNT cell-transferred mice. ( B ) Representative H&E staining and Oil Red O and α-SMA staining in livers from NCD- and MCD-fed mice and TCRαβ + DNT-transferred mice. ( C ) NAFLD activity score (NAS) in livers of NCD- and MCD-fed mice and TCRαβ + DNT-transferred mice. ( D–F ) Representative flow cytometry plots and statistical analysis of proportion of CD4 + and CD8 + Treg cells among livers of NCD- and MCD-fed mice and TCRαβ + DNT cell-transferred mice. ( G ) Representative flow cytometry plots and statistical analysis of apoptosis of intrahepatic CD4 + and CD8 + cells of NCD- and MCD-fed mice and TCRαβ + DNT cell-transferred mice. n ≥ 4 mice/group. Two-sided P values <.05 were considered significant. ∗∗ P < .01; ∗ P < .05.

Article Snippet: After being allowed to adapt for 5 days, the C57BL/6 mice were fed either a NCD or a MCD (10401; Beijing HFK Bioscience, Beijing, China) for 5 weeks or a CDHFD for 18 weeks.

Techniques: Adoptive Transfer Assay, Staining, Activity Assay, Flow Cytometry

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Critical and Diverse Roles of CD4 – CD8 – Double Negative T Cells in Nonalcoholic Fatty Liver Disease

doi: 10.1016/j.jcmgh.2022.02.019

Figure Lengend Snippet: FAs are involved in regulating apoptosis and the function of TCR subtypes of DNT cells. ( A ) GSEA of the response to FA metabolism. ( B ) The level of Bodipy in intrahepatic TCRαβ + DNT cells and TCRγδ + DNT cells in NCD- and MCD-fed mice. n = 6 mice/group. ( C ) CD36 expression in intrahepatic DNT cells of NCD- and MCD-fed mice by flow cytometry and real-time PCR. n = 6 mice/group. ( D ) Principal component analysis of metabonomics in livers of NCD- and MCD-fed mice. ( E ) FA content in livers of NCD- and MCD-fed mice. n = 6 mice/group. ( F ) Correlation between the harmful FAs ADA, AA levels, and DNT cell apoptosis. n = 9 mice/group. ( G ) Apoptosis, Bodipy, GZMB, IL17A expression in TCRγδ + and TCRαβ + DNT cells after ADA and AA stimulation. n = 6 biologically independent samples per group. Two-sided P values <.05 were considered significant. ∗∗ P < .01; ∗ P < .05.

Article Snippet: After being allowed to adapt for 5 days, the C57BL/6 mice were fed either a NCD or a MCD (10401; Beijing HFK Bioscience, Beijing, China) for 5 weeks or a CDHFD for 18 weeks.

Techniques: Expressing, Flow Cytometry, Real-time Polymerase Chain Reaction

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Critical and Diverse Roles of CD4 – CD8 – Double Negative T Cells in Nonalcoholic Fatty Liver Disease

doi: 10.1016/j.jcmgh.2022.02.019

Figure Lengend Snippet: The variation tendency of DNT cells in livers of NAFLD patients was consistent with that in mice. ( A ) Typical picture of multiplex immunofluorescence staining of human liver sections. TCRαβ ( green ), CD4/CD8/CD56 ( red ). White arrowheads indicate DNT cells. Scale bar, 25 μm. ( B ) Number of TCRαβ + DNT cells per area in human liver. The number of DNT cells per area (150 μm × 150 μm) was counted. ( C ) Absolute number of TCRαβ + DNT cells per g mouse liver. ( D ) Typical picture of multiplex immunofluorescence staining of human liver sections. TCRγδ ( green ), CD4/CD8/CD56 ( red ). White arrowheads indicate DNT cells. Scale bar, 25 μm. ( E ) Number of TCRαβ + DNT cells per area in human liver. The number of DNT cells per area (150 μm × 150 μm) was counted. ( F ) Absolute number of TCRγδ + DNT cells per g mouse liver. ( G ) The amplitude of number variation of TCRγδ + CD4 – CD8 – CD56 – T cells and TCRαβ + CD4 – CD8 – CD56 – T cells in NAFLD/Health. ( H ) The amplitude of number variation of TCRαβ + DNT cells and TCRγδ + DNT cells in MCD/NCD. ( I ) Ratio of TCRαβ + DNT cells with CD4 + CD8 + CD56 + cells (0.02 cm 2 per area). ( J ) Ratio of TCRγδ + DNT cells with CD4 + CD8 + CD56 + cells (0.02 cm 2 per area). Two-sided P values <.05 were considered significant. ∗∗ P < .01; ∗ P < .05.

Article Snippet: After being allowed to adapt for 5 days, the C57BL/6 mice were fed either a NCD or a MCD (10401; Beijing HFK Bioscience, Beijing, China) for 5 weeks or a CDHFD for 18 weeks.

Techniques: Multiplex Assay, Immunofluorescence, Staining

Journal:

Article Title: Liver-Specific ?-Catenin Knockout Mice Exhibit Defective Bile Acid and Cholesterol Homeostasis and Increased Susceptibility to Diet-Induced Steatohepatitis

doi: 10.2353/ajpath.2010.090667

Figure Lengend Snippet: β-Catenin knockout mice fed the MCD diet develop severe steatohepatitis. A: Gross, H&E-, and oil red O–stained sections of livers from wild-type or KO mice fed either the control or MCD diets as indicated (liver sections at ×200 magnification). B: Quantification of steatosis, inflammation, and hepatocyte ballooning seen in the H&E stained sections by a pathologist blinded to the treatment groups. Values are expressed as means ± SEM from five to seven animals per group. *P < 0.05 wild-type versus KO groups, **P < 0.05 CTL versus MCD groups. WT/CTL indicates wild-type mice on control diet; WT/MCD, wild-type mice on MCD diet; KO/CTL, KO mice on the control diet; KO/MCD, KO mice on MCD diet.

Article Snippet: Mice were fed the MCD diet or the corresponding control diet, which was supplemented with 3 g/kg methionine and 2 g/kg choline (ICN Biomedicals, Costa Mesa, CA, catalog numbers 960439 and 960441, respectively).

Techniques: Knock-Out, Staining, Control

Journal:

Article Title: Liver-Specific ?-Catenin Knockout Mice Exhibit Defective Bile Acid and Cholesterol Homeostasis and Increased Susceptibility to Diet-Induced Steatohepatitis

doi: 10.2353/ajpath.2010.090667

Figure Lengend Snippet: β-Catenin knockout mice on the MCD diet have more liver injury and fibrosis. A: Serum ALT and AST levels. B, Upper panel: Representative sections of livers from the four experimental groups stained with Trichrome and Reticulin stain (×200 magnification). Enhanced fibrosis is visible as blue color in Trichrome staining (×400; inset). Coarse black reticulin fibers consistent with increased pericellular fibrosis and focal early bridging can be seen in the KO/MCD group. B, Lower panel: Representative liver sections stained with α-SMA. The white arrowhead indicates increased α-SMA staining (brown color) in the KO/MCD group. C: Quantification of fibrosis and pericellular fibrosis of trichrome stained sections by a pathologist blinded to the treatment groups. Results are expressed as means ± SEM of samples from at least five animals, **P < 0.05 compared with all other groups, *P < 0.05 wild-type versus KO groups, ***P < 0.05 CTL versus MCD groups. WT/CTL indicates wild-type mice on control diet; WT/MCD, wild-type mice on MCD diet; KO/CTL, knockout mice on the control diet; KO/MCD, knockout mice on MCD diet.

Article Snippet: Mice were fed the MCD diet or the corresponding control diet, which was supplemented with 3 g/kg methionine and 2 g/kg choline (ICN Biomedicals, Costa Mesa, CA, catalog numbers 960439 and 960441, respectively).

Techniques: Knock-Out, Staining, Control

Journal:

Article Title: Liver-Specific ?-Catenin Knockout Mice Exhibit Defective Bile Acid and Cholesterol Homeostasis and Increased Susceptibility to Diet-Induced Steatohepatitis

doi: 10.2353/ajpath.2010.090667

Figure Lengend Snippet: Levels of β-catenin and its target genes in knockout and wild-type animals fed the control or MCD diets. A, Upper panel: Immunohistochemical staining with total β-catenin. Brown staining for β-catenin is visible in the two wild-type groups at the cell membrane. No staining is seen in the two KO groups. A, Lower panel: Glutamine synthetase staining. Brown glutamine synthetase staining is visible in a narrow rim of perivenous hepatocytes from wild-type animals fed either the control or MCD diets (black arrowhead). No staining is visible in KO livers (All images are at ×200 magnification). B: Western blot analysis for total β-catenin using liver lysates from the four experimental groups. Two representative samples from each experimental group are shown. The membrane was stripped and reprobed with β-actin as internal loading control. C: Western blot analysis for Cyp2E1 using liver microsomal preparation from each of the experimental groups. One representative sample from each group is shown. The membrane was stained with Ponceau red to confirm equal loading. WT/CTL indicates wild-type mice on control diet; WT/MCD, wild-type mice on MCD diet; KO/CTL, knockout mice on the control diet; KO/MCD, knockout mice on MCD diet.

Article Snippet: Mice were fed the MCD diet or the corresponding control diet, which was supplemented with 3 g/kg methionine and 2 g/kg choline (ICN Biomedicals, Costa Mesa, CA, catalog numbers 960439 and 960441, respectively).

Techniques: Knock-Out, Control, Immunohistochemical staining, Staining, Membrane, Western Blot

Journal:

Article Title: Liver-Specific ?-Catenin Knockout Mice Exhibit Defective Bile Acid and Cholesterol Homeostasis and Increased Susceptibility to Diet-Induced Steatohepatitis

doi: 10.2353/ajpath.2010.090667

Figure Lengend Snippet: Hepatic triglyceride and cholesterol levels in control-diet and MCD-diet fed wild-type and KO mice. A: Hepatic triglyceride levels. B: Hepatic cholesterol levels. Results are expressed as means ± SEM of five samples per group. *P < 0.05 wild-type versus KO groups, *P < 0.05 CTL versus MCD groups. WT/CTL indicates wild-type mice on control diet; WT/MCD, wild-type mice on MCD diet; KO/CTL, knockout mice on the control diet; KO/MCD, knockout mice on MCD diet.

Article Snippet: Mice were fed the MCD diet or the corresponding control diet, which was supplemented with 3 g/kg methionine and 2 g/kg choline (ICN Biomedicals, Costa Mesa, CA, catalog numbers 960439 and 960441, respectively).

Techniques: Control, Knock-Out

Journal:

Article Title: Liver-Specific ?-Catenin Knockout Mice Exhibit Defective Bile Acid and Cholesterol Homeostasis and Increased Susceptibility to Diet-Induced Steatohepatitis

doi: 10.2353/ajpath.2010.090667

Figure Lengend Snippet: Expression analysis by quantitative real-time PCR of genes involved in hepatic fatty acid and triglyceride metabolism. Results are expressed as means ± SEM of five to seven samples per group, with each sample analyzed in duplicate. *P < 0.05 wild-type versus KO groups, **P < 0.05 CTL versus MCD groups. WT/CTL indicates wild-type mice on control diet; WT/MCD, wild-type mice on MCD diet; KO/CTL, knockout mice on the control diet; KO/MCD, knockout mice on MCD diet.

Article Snippet: Mice were fed the MCD diet or the corresponding control diet, which was supplemented with 3 g/kg methionine and 2 g/kg choline (ICN Biomedicals, Costa Mesa, CA, catalog numbers 960439 and 960441, respectively).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Knock-Out

Journal:

Article Title: Liver-Specific ?-Catenin Knockout Mice Exhibit Defective Bile Acid and Cholesterol Homeostasis and Increased Susceptibility to Diet-Induced Steatohepatitis

doi: 10.2353/ajpath.2010.090667

Figure Lengend Snippet: Expression analysis by quantitative real-time PCR of genes involved in hepatic cholesterol metabolism. A: Genes involved in cholesterol biosynthesis. B: Genes involved in cholesterol efflux and transport. C: Genes encoding the cytochrome P450 enzymes involved in the metabolism of cholesterol to bile acids. Results are expressed as means ± SEM of five to seven samples per group, with each sample analyzed in duplicate. *P < 0.05 wild-type versus KO groups, **P < 0.05 CTL versus MCD groups. WT/CTL indicates wild-type mice on control diet; WT/MCD, wild-type mice on MCD diet; KO/CTL, knockout mice on the control diet; KO/MCD, knockout mice on MCD diet.

Article Snippet: Mice were fed the MCD diet or the corresponding control diet, which was supplemented with 3 g/kg methionine and 2 g/kg choline (ICN Biomedicals, Costa Mesa, CA, catalog numbers 960439 and 960441, respectively).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Knock-Out

Journal:

Article Title: Liver-Specific ?-Catenin Knockout Mice Exhibit Defective Bile Acid and Cholesterol Homeostasis and Increased Susceptibility to Diet-Induced Steatohepatitis

doi: 10.2353/ajpath.2010.090667

Figure Lengend Snippet: Analysis of hepatic bile acids and serum bilirubin levels. A: Hepatic total bile acid levels in wild-type and KO mice on control or MCD diet. B: Analysis of expression levels by real-time PCR of genes involved in export of bile acids and bilirubin. Data are expressed as arbitrary units relative to the expression level in the WT/CTL group. C: Representative serum samples from each treatment group. D: Serum total bilirubin levels. E: Analysis of expression levels of Hmox1 by real-time PCR. Results are expressed as means ± SEM of 5 samples per group, with each sample analyzed in duplicate. **P < 0.01 compared with all other groups, *P < 0.05 wild-type versus KO groups. WT/CTL indicates wild-type mice on control diet; WT/MCD, wild-type mice on MCD diet; KO/CTL, knockout mice on the control diet; KO/MCD, knockout mice on MCD diet.

Article Snippet: Mice were fed the MCD diet or the corresponding control diet, which was supplemented with 3 g/kg methionine and 2 g/kg choline (ICN Biomedicals, Costa Mesa, CA, catalog numbers 960439 and 960441, respectively).

Techniques: Control, Expressing, Real-time Polymerase Chain Reaction, Knock-Out

Journal: The Journal of Clinical Investigation

Article Title: Oxidative stress promotes pathologic polyploidization in nonalcoholic fatty liver disease

doi: 10.1172/JCI73957

Figure Lengend Snippet: (A) MCD diet model. Hepatocytes were separated into ploidy populations by FACS analysis (n = 3 per group) with 2c, 4c, and 8c and >8c DNA content corresponding to diploid, tetraploid, and highly polyploid hepatocytes, respectively. Images of liver sections after double staining with anti–β-catenin (plasma membrane labeling, red) and Hoechst (nucleus, green) in WT mice fed control (CTRL) or MCD diets (scale bar: 20 μm). Box plots of the percentage of ≥8n mononuclear hepatocytes relative to total hepatocytes in control-fed and MCD diet–fed mice. Results represent mean ± SEM (n = 8 per group). ***P < 0.001, Student’s t test. (B) HFD model. Hepatocytes were separated into ploidy populations by FACS analysis (n = 3 per group) with 2c, 4c, and 8c and >8c DNA content corresponding to diploid, tetraploid, and highly polyploid hepatocytes, respectively. Images of liver sections after double staining with anti–β-catenin (red) and Hoechst (green) in WT mice fed a control or a HFD (scale bar: 20 μm). Box plots of the percentage of ≥8n mononuclear hepatocytes relative to total hepatocytes in control-fed and HFD-fed mice. Results represent mean ± SEM (n = 5 per group). ***P < 0.001, Student’s t test.

Article Snippet: For MCD diet–induced fatty liver, 10-week-old mice were fed for 3 to 5 weeks with a MCD diet (Ssniff EF R/M, MCD mod) purchased from Ssniff Spezialdiäten GmbH.

Techniques: Double Staining, Clinical Proteomics, Membrane, Labeling, Control

Journal: The Journal of Clinical Investigation

Article Title: Oxidative stress promotes pathologic polyploidization in nonalcoholic fatty liver disease

doi: 10.1172/JCI73957

Figure Lengend Snippet: (A) Schematic representation of the nuclei double-labeling experiment with thymidine analogs. C57BL/6J mice were fed MCD diet for 4 weeks (d1–d27). At d15, animals had access to labeled CldU drinking water for 5 days. Over following 2 days, animals had access to unlabeled water to clear unincorporated CldU, followed by 5 days of access to IdU drinking water. (B) Images of hepatocyte nuclei from MCD diet liver sections after costaining with anti-CldU (green) and anti-IdU (red) and a merge with Hoechst (blue) (scale bar: 50 μm). The arrowheads point to double-labeled nuclei (yellow) (scale bar: 200 μm). (C) Quantification of polyploid mononuclear fatty hepatocytes with nuclei double labeling (CldU+IdU+) in MCD diet liver (n = 4). Results represent mean ± SEM.

Article Snippet: For MCD diet–induced fatty liver, 10-week-old mice were fed for 3 to 5 weeks with a MCD diet (Ssniff EF R/M, MCD mod) purchased from Ssniff Spezialdiäten GmbH.

Techniques: Labeling