Journal: Nutrients

Article Title: Metabolic Health in Obese Subjects—Is There a Link to Lactoferrin and Lactoferrin Receptor-Related Gene Polymorphisms?

doi: 10.3390/nu12092843

Figure Lengend Snippet: Distribution of alleles and genotypes in analysed gene polymorphisms in MHO and MUHO subjects.

Article Snippet: SNPs in LTF rs1126477, LTF rs2239692, LTF rs1126478, low-density lipoprotein receptor-related protein 2 ( LRP2 ) rs2544390, LRP1 rs4759277, and LRP1 rs1799986, genes were genotyped with TaqMan allelic discrimination assays (Applied Biosystem, Foster City, NC, USA): rs2544390: C___8822318_10, rs4759277: C___31186847_10, rs1799986: C___1955081_10, rs1126477: C___9698511_10; rs2239692: C___2610649_10; rs1126478: C___9698521_10) using Bio-Rad CFX96TM Real-Time PCR system (Hercules, CA, USA).

Techniques:

Journal: Nutrients

Article Title: Metabolic Health in Obese Subjects—Is There a Link to Lactoferrin and Lactoferrin Receptor-Related Gene Polymorphisms?

doi: 10.3390/nu12092843

Figure Lengend Snippet: Odds ratios (OR) and 95% confidence interval (CI) for the associations between analysed gene polymorphisms and the prevalence of metabolic disorders in obese subjects.

Article Snippet: SNPs in LTF rs1126477, LTF rs2239692, LTF rs1126478, low-density lipoprotein receptor-related protein 2 ( LRP2 ) rs2544390, LRP1 rs4759277, and LRP1 rs1799986, genes were genotyped with TaqMan allelic discrimination assays (Applied Biosystem, Foster City, NC, USA): rs2544390: C___8822318_10, rs4759277: C___31186847_10, rs1799986: C___1955081_10, rs1126477: C___9698511_10; rs2239692: C___2610649_10; rs1126478: C___9698521_10) using Bio-Rad CFX96TM Real-Time PCR system (Hercules, CA, USA).

Techniques:

Journal: Disease Models & Mechanisms

Article Title: Alterations in nuclear structure promote lupus autoimmunity in a mouse model

doi: 10.1242/dmm.024851

Figure Lengend Snippet: Anti-neutrophil serum reactivity develops in (NZW×B6.Lbr ic )F 1 mice. (A) Fluorescence microscopy of ethanol-fixed human PMNs stained with DAPI (top panel), and female (NZW×B6.Lbr ic )F 1 mouse serum and an Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (bottom panel). Broken circles indicate DAPI-positive bilobed eosinophils that were not readily detected by the female (NZW×B6.Lbr ic )F 1 mouse serum. Image magnification, 100×. (B) High-power magnification of ethanol-fixed PMN staining. Neutrophils displayed strong nuclear staining, but eosinophils had comparatively weak and patchy perinuclear staining. Image magnification, 400×. The preferential staining of isolated neutrophils relative to peripheral blood mononuclear cells and isolated monocytes is demonstrated in greater detail in Fig. S7 . (C) Ethanol-fixed PMNs were co-stained with female (NZW×B6.Lbr ic )F 1 mouse serum (green) and anti-MPO (red), nuclei were counterstained with DAPI (blue). The mouse serum staining did not colocalize with MPO. Image magnification, 1000×.

Article Snippet: HEp-2 cells immobilized on microscopy slides (MBL/Bion) were incubated with a 1:200 dilution of mouse sera, and a FITC-conjugated goat anti-mouse immunoglobulin secondary antibody (1:1000; 1010-02, Southern Biotech).

Techniques: Fluorescence, Microscopy, Staining, Isolation

Journal: PLOS Pathogens

Article Title: Isolation and characterization of IgG3 glycan-targeting antibodies with exceptional cross-reactivity for diverse viral families

doi: 10.1371/journal.ppat.1012499

Figure Lengend Snippet: (A) HEp-2 cell slides were immunofluorescently stained with the indicated mAbs and goat anti-human Ig-FITC secondary (green) and DAPI (blue, to stain cell nuclei). Each mAb was tested at 100, 10, and 1 μg/mL. Control human serum samples that were positive or negative for anti-nuclear antibodies are shown (right). Images are 40X magnification. (B) Autoantigens were tested for reactivity by 2526, Ab82 (negative control antibody), and A32 (positive control antibody) using a commercially available AtheNA Multi-Lyte ANA kit. Values shown in bold were considered positive.

Article Snippet: HEp-2 cell coated slides (BION ENTERPRISES LTD ANA (HEp-2) Test System, ANK-120) were incubated with purified antibodies at 100, 10, or 1 μg/ml or control sera in a moist chamber at room temperature for 30 min. Controls provided with the kit included anti-nuclear antibody (ANA)+ and ANA- human sera.

Techniques: Staining, Control, Negative Control, Positive Control

Journal: Frontiers in Immunology

Article Title: Upregulated Fcrl5 disrupts B cell anergy causes autoimmune disease

doi: 10.3389/fimmu.2023.1276014

Figure Lengend Snippet: Aged Fcrl5 Tg mice develop autoimmune disease. (A) Representative images of ANA staining obtained with serum from young WT (n=12), young Fcrl5 Tg (n=13), aged WT (n=19), and aged Fcrl5 Tg (n=17) mice, detected by immunofluorescence assay using HEp-2 cells. Scale Bars, 50 μm. Data are pooled from three independent experiments. (B) Autoantibody production against dsDNA, histone, and Sm/RNP (ribonucleoprotein) in serum isolated from young WT (n=11), young Fcrl5 Tg (n=12), aged WT (n=6), and aged Fcrl5 Tg (n=11) mice, assayed by ELISA. OD, optical density. Data are pooled from three independent experiments. (C) Left, representative H&E-stained histological lung, liver, and kidney images of young WT (n=6), young Fcrl5 Tg (n=8), aged WT (n=6), and aged Fcrl5 Tg (n=14) mice. Arrowheads indicate areas of cell infiltration. Scale bars, 200 μm. Data are representative of three independent experiments. Right, quantitated cell infiltration (cell infiltration area per total area) in the lung, liver, and kidney and mean linear intercept (MLI) in the lung of young WT (n=6), young Fcrl5 Tg (n=8), aged WT (n=6), and aged Fcrl5 Tg (n=14) mice. Data are pooled from three independent experiments. Statistical data are shown as mean values with s.d., and data were analyzed by one-way ANOVA with Tukey’s multiple comparisons test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: Diluted mouse serum (1:100 in 0.1% BSA in PBS) was incubated on HEp-2 slides (MBL or Bio-Rad) for 1 h in the dark at room temperature.

Techniques: Staining, Immunofluorescence, Isolation, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Upregulated Fcrl5 disrupts B cell anergy causes autoimmune disease

doi: 10.3389/fimmu.2023.1276014

Figure Lengend Snippet: Upregulation of Fcrl5 in B cells exacerbates SLE-like disease model. (A) A scheme for induction of an imiquimod (IMQ)-induced SLE-like model. (B) Representative images of ANA staining obtained with serum from imiquimod-untreated (IMQ–) or -treated (IMQ+) WT (n=16) and Fcrl5 Tg (n=18) mice, detected by immunofluorescence assay using HEp-2 cells. Scale Bars, 50 μm. Data are pooled from three independent experiments. (C) Autoantibody production against dsDNA, histone, and Sm/RNP (ribonucleoprotein) in serum isolated from imiquimod-untreated (IMQ–) or -treated (IMQ+) WT (n=12) and Fcrl5 Tg (n=14) mice, assayed by ELISA. OD, optical density. Data are pooled from three independent experiments. (D) Representative H&E-stained histological kidney images and quantitated cell infiltration (cell infiltration area per total area) in the kidney of imiquimod-untreated (IMQ–) WT (n=9) and Fcrl5 Tg (n=9) or -treated (IMQ+) WT (n=14) and Fcrl5 Tg (n=16) mice. Arrowheads indicate areas of cell infiltration. Scale bars, 200 μm. Data are pooled from three independent experiments. Statistical data are shown as mean values with s.d., and data were analyzed by one-way ANOVA with Tukey’s multiple comparisons test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: Diluted mouse serum (1:100 in 0.1% BSA in PBS) was incubated on HEp-2 slides (MBL or Bio-Rad) for 1 h in the dark at room temperature.

Techniques: Staining, Immunofluorescence, Isolation, Enzyme-linked Immunosorbent Assay