Journal: Scientific Reports

Article Title: Dietary wheat amylase trypsin inhibitors promote features of murine non-alcoholic fatty liver disease

doi: 10.1038/s41598-019-53323-x

Figure Lengend Snippet: ATI feeding increases liver macrophage numbers and their M1- vs M2-type polarization. ( A – C ) Immunohistochemistry and quantitative morphometry for CD68 and YM-1 positive cells (original magnification 40x). ( D ) Ratio of total (CD68+) vs M2-type (Ym-1+) macrophages. ( E ) CD11b+ F4/80+ macrophage subset (% of CD45 positive total immune cells) as determined by FACS analysis. Comparisons by ANOVA; data are means ± SEM for 10 representative sections per mouse and 7–10 mice per group; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Tissue was blocked with 5% normal donkey serum, and subsequently incubated with primary antibodies of to CD68 (1:100, Biozol, clone: FA-11), CD86 (1:100, Abcam, cat no: ab119857), and MHC-II (1:100, Abcam, Cat no: 180779) for 2 hrs at room temperature and finally incubated with respective Alexa-flour 488 labelled secondary antibodies.

Techniques: Immunohistochemistry

Journal: Scientific Reports

Article Title: Dietary wheat amylase trypsin inhibitors promote features of murine non-alcoholic fatty liver disease

doi: 10.1038/s41598-019-53323-x

Figure Lengend Snippet: ATI feeding increases hepatic pro-inflammatory and macrophage M1- vs M2-type gene expression. ( A – F ) Hepatic transcript levels of cd68, tnfa, il1b, il6, arg1 and ym1. Comparisons by ANOVA; data are expressed as means ± SEM for 7–10 mice per group; *p < 0.05, **p < 0.01.

Article Snippet: Tissue was blocked with 5% normal donkey serum, and subsequently incubated with primary antibodies of to CD68 (1:100, Biozol, clone: FA-11), CD86 (1:100, Abcam, cat no: ab119857), and MHC-II (1:100, Abcam, Cat no: 180779) for 2 hrs at room temperature and finally incubated with respective Alexa-flour 488 labelled secondary antibodies.

Techniques: Gene Expression

Journal: Scientific Reports

Article Title: Dietary wheat amylase trypsin inhibitors promote features of murine non-alcoholic fatty liver disease

doi: 10.1038/s41598-019-53323-x

Figure Lengend Snippet: Nutritional ATI promote central adipose tissue inflammation. ( A ) Crown like structures (CLS = accumulation of macrophages) in CD68+ stained sections of epididymal adipose tissue in the 4 experimental groups (original magnification 40x), the number of CD68+ CLS as determined by morphometry, and epididymal fat as % of body weight. ( B ) fat weights, and ( C ) transcript levels of cd68, il6 and il1b. Comparisons by ANOVA; data are means ± SEM for 10 representative sections per mouse and 7–10 mice per group; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Tissue was blocked with 5% normal donkey serum, and subsequently incubated with primary antibodies of to CD68 (1:100, Biozol, clone: FA-11), CD86 (1:100, Abcam, cat no: ab119857), and MHC-II (1:100, Abcam, Cat no: 180779) for 2 hrs at room temperature and finally incubated with respective Alexa-flour 488 labelled secondary antibodies.

Techniques: Staining

Journal: Scientific Reports

Article Title: Dietary wheat amylase trypsin inhibitors promote features of murine non-alcoholic fatty liver disease

doi: 10.1038/s41598-019-53323-x

Figure Lengend Snippet: ATI feeding increases intestinal macrophage and dendritic cell activation and maturation. ( A–C ) CD68, CD86 and MCH-II expressing cells in the terminal ileum; scale bar: 100 and 50 µm. ( D ) Morphometric quantification of CD68, CD86 and MHC-II positive cells. ( E ) Transcript levels of il1b, tnfα and il6. Comparisons by ANOVA; data are expressed as means ± SEM of 6 mice per group and 5 representative sections per mouse; *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Tissue was blocked with 5% normal donkey serum, and subsequently incubated with primary antibodies of to CD68 (1:100, Biozol, clone: FA-11), CD86 (1:100, Abcam, cat no: ab119857), and MHC-II (1:100, Abcam, Cat no: 180779) for 2 hrs at room temperature and finally incubated with respective Alexa-flour 488 labelled secondary antibodies.

Techniques: Activation Assay, Expressing

Journal: Frontiers in Immunology

Article Title: The C1q-ApoE complex: A new hallmark pathology of viral hepatitis and nonalcoholic fatty liver disease

doi: 10.3389/fimmu.2022.970938

Figure Lengend Snippet: C1q- and ApoE-expressing cells in 3 month old WT and Mdr2-KO mouse livers. (A) Liver sections were stained with C1q (white), Clec4f (red) for KCs, CD68 (green) for macrophages and DAPI (blue) for nuclei. Dotted lines demarcate immune cell infiltrates in the portal tract areas from surrounding liver tissue. KCs are known to stain CD68 + /Clec4f + . (B) Liver sections were stained with ApoE (white), Clec4f (red) for KCs, CD68 (green) for macrophages and DAPI (blue) for nuclei. Dotted lines demarcate immune cell infiltrates in the portal tract areas from surrounding liver tissue. Representative images of WT livers (n=6); Mdr2-KO livers (n=6) are shown. Scale bars 100 μm.

Article Snippet: Immunoflorescent (IF) stainings of mouse liver sections were performed using the following primary antibodies: anti-mouse/human ApoE (ab52607; Abcam); anti-mouse C1q (ab182451; Abcam); anti-mouse CLEC4F (ab2608299; Invitrogen); anti-mouse CD68 (FA11; Serotec); anti-mouse CD31 (ab553370; BD PharMingen); anti-human C3 (A213; ComplementTech); anti-mouse C4 (HM1046; Hycult Biotech); anti-mouse C5 (ab11898, Abcam).

Techniques: Expressing, Staining

Journal: Frontiers in Immunology

Article Title: The C1q-ApoE complex: A new hallmark pathology of viral hepatitis and nonalcoholic fatty liver disease

doi: 10.3389/fimmu.2022.970938

Figure Lengend Snippet: Complement component C4 and C5 expression in WT and Mdr2-KO mouse livers. (A) Liver sections were stained with Clec4f (green) for KCs, C4 (red), and DAPI for nuclei (blue). Dotted lines demarcate Clecf + cells as well as immune cell infiltrates in portal triads from adjacent liver parenchyma. (B) Liver sections were stained with Clec4f (green) for KCs, C5 (red), and DAPI (blue) for nuclei. (C) Complement component C3, C1q, CD68 expression in WT and Mdr2-KO livers. Liver sections were stained with C3 (red), C1q (white), and CD68 (green) for macrophages/KCs and DAPI for nuclei (blue). Dotted lines demarcate immune cell infiltration sites from adjacent liver parenchyma. White squares represent high magnification images shown separately. Representative images of WT livers (n=6); Mdr2-KO livers (n=6) are shown. Scale bars 100 μm in images of low and high magnification.

Article Snippet: Immunoflorescent (IF) stainings of mouse liver sections were performed using the following primary antibodies: anti-mouse/human ApoE (ab52607; Abcam); anti-mouse C1q (ab182451; Abcam); anti-mouse CLEC4F (ab2608299; Invitrogen); anti-mouse CD68 (FA11; Serotec); anti-mouse CD31 (ab553370; BD PharMingen); anti-human C3 (A213; ComplementTech); anti-mouse C4 (HM1046; Hycult Biotech); anti-mouse C5 (ab11898, Abcam).

Techniques: Expressing, Staining

Journal: Frontiers in Immunology

Article Title: The C1q-ApoE complex: A new hallmark pathology of viral hepatitis and nonalcoholic fatty liver disease

doi: 10.3389/fimmu.2022.970938

Figure Lengend Snippet: Translational studies in healthy and diseased human livers. (A) Normal liver sample Nr. 5 ( Supplementary Table 1 ), NAFLD sample Nr. 9 ( Supplementary Table 1 ) and viral hepatitis sample Nr. 7 ( Supplementary Table 1 ) tissues were stained with ORO for lipid (red) and HE for nuclei (blue). (B) Human livers - as described in Figure 6A - were stained with ApoE (red), C1q (white), and DAPI (blue) showing similar staining patterns vs mouse livers. Dotted lines demarcate immune cell infiltration sites from adjacent liver parenchyma. (C) Human livers – as described in Figure 6A - were stained with C5 (red), CD68 (green) and DAPI (blue) indicating nuclei. Representative images of human samples ( Supplementary Table 1 ) are shown. Dotted lines demarcate immune cell infiltration sites from adjacent liver parenchyma. Scale bars 100 μm.

Article Snippet: Immunoflorescent (IF) stainings of mouse liver sections were performed using the following primary antibodies: anti-mouse/human ApoE (ab52607; Abcam); anti-mouse C1q (ab182451; Abcam); anti-mouse CLEC4F (ab2608299; Invitrogen); anti-mouse CD68 (FA11; Serotec); anti-mouse CD31 (ab553370; BD PharMingen); anti-human C3 (A213; ComplementTech); anti-mouse C4 (HM1046; Hycult Biotech); anti-mouse C5 (ab11898, Abcam).

Techniques: Staining

Journal:

Article Title: Activated microglia in cortex of mouse models of mucopolysaccharidoses I and IIIB

doi: 10.1073/pnas.252784899

Figure Lengend Snippet: Histochemical and immunohistochemical characterization of microglia in cortex. (A) G. simplicifolia isolectin IB4-positive microglia apposed to neuron; MPS IIIB, 7 months (×1,750). (B) MOMA-2-positive microglia apposed to neuron; MPS IIIB, 3 months (×1,150). (C and D) MOMA-2-positive cells seen at low power; MPS IIIB 3 months and MPS I, 14 months, respectively (×140). (E–G) Confocal images of sections stained with antibody against Lamp-1 (green), MOMA-2 (red), and merged; MPS IIIB, 6 months (×350). (H–J) Fluorescent images using antibody against ganglioside GM3 (green), MOMA-2 (red), and merged; MPS IIIB, 6 months (×260). (K–M) Images of sections stained with antibody against CD68/macrosialin, MPS IIIB, MPS I, and control, respectively, 3 months (×65). (N and O) Images of sections stained with antibody against IFN-γ; MPS IIIB, and control, respectively, 3 months (×250). (P and Q) Images of sections stained with antibody against IFN-γ receptor; MPS IIIB and control, respectively, 7 months (×250).

Article Snippet: A rat monoclonal antibody against mouse macrophages/monocytes (MOMA-2) and a rat monoclonal antibody against mouse CD68 (macrosialin) were purchased from Serotec.

Techniques: Immunohistochemical staining, Staining

Journal:

Article Title: Activated microglia in cortex of mouse models of mucopolysaccharidoses I and IIIB

doi: 10.1073/pnas.252784899

Figure Lengend Snippet: Transcripts increased in cortex of mouse models of MPS IIIB and MPS I, determined by microarray analysis

Article Snippet: A rat monoclonal antibody against mouse macrophages/monocytes (MOMA-2) and a rat monoclonal antibody against mouse CD68 (macrosialin) were purchased from Serotec.

Techniques: Microarray, Binding Assay

Journal:

Article Title: Activated microglia in cortex of mouse models of mucopolysaccharidoses I and IIIB

doi: 10.1073/pnas.252784899

Figure Lengend Snippet: Increase in transcripts in cortex of mice at different ages determined by Northern blot analysis

Article Snippet: A rat monoclonal antibody against mouse macrophages/monocytes (MOMA-2) and a rat monoclonal antibody against mouse CD68 (macrosialin) were purchased from Serotec.

Techniques: Northern Blot