Journal: Frontiers in Immunology

Article Title: Circulating and Salivary Antibodies to Fusobacterium nucleatum Are Associated With Cystic Pancreatic Neoplasm Malignancy

doi: 10.3389/fimmu.2020.02003

Figure Lengend Snippet: Circulating plasma antibody reactivity to oral commensals identified in pancreatic tumor lesions. Plasma diluent (1/300) from patients with histology verified pancreas tissues: IPMN with low grade dysplasia (LGD-IPMN), IPMN with high grade dysplasia or with invasive cancer (HGD-IPMN + Cancer), or benign tumors/non-IPMN (Control) was tested in duplicates in ELISA plates coated with indicated antigen in (A–F) (bacteria: 5 × 10E7 CFU/ml, peptide 10 μg/mL). The total plasma IgG level is shown in (G) . Internal control Mean OD absorbance values of duplicates are shown. Statistical analysis was performed using the Kolmogorov–Smirnov test, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Total levels of salivary IgA and plasma IgG antibodies were measured using Human IgA/IgG ELISA Kit (Novus Biologicals, Colorado, United States) according to the manufacturer’s instructions.

Techniques: Clinical Proteomics, Control, Enzyme-linked Immunosorbent Assay, Bacteria

Journal: Frontiers in Immunology

Article Title: Circulating and Salivary Antibodies to Fusobacterium nucleatum Are Associated With Cystic Pancreatic Neoplasm Malignancy

doi: 10.3389/fimmu.2020.02003

Figure Lengend Snippet: Salivary IgA antibody reactivity to oral commensals quantified by ELISA using saliva diluent (1/16) from indicated patient group. Salivary IgA reactivity to F. nucleatum , the Fap2 mimotope, and S. gordonii (A,B,F) in HG-IPMN/Cancer patients is significantly increased compared to other groups. The differences in antibody levels against bacterial antigens of G. adiacens, S. anginosus , and E. coli (C,D,E) were insignificant between groups. Total plasma IgA level did not differ between the groups (G) . Statistical analysis was performed using the Kolmogorov–Smirnov test, * p < 0.05 and ** p < 0.01.

Article Snippet: Total levels of salivary IgA and plasma IgG antibodies were measured using Human IgA/IgG ELISA Kit (Novus Biologicals, Colorado, United States) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: Frontiers in Immunology

Article Title: Circulating and Salivary Antibodies to Fusobacterium nucleatum Are Associated With Cystic Pancreatic Neoplasm Malignancy

doi: 10.3389/fimmu.2020.02003

Figure Lengend Snippet: Correlation between circulating plasma (A) or salivary (B) antibody reactivity to F. nucleatum and Fap2 mimotope, or between circulating plasma antibody and salivary antibodies to either of antigen (C,D) . Competitive ELISA (E) with saliva samples with or without antigen pre-absorption as indicated for 2 h, then subjected to Fap2 mimotope ELISA test. Statistical analysis was performed using two-tailed Pearson correlation test and Wilcoxon test for two related samples. ** p < 0.01, ns = not significant.

Article Snippet: Total levels of salivary IgA and plasma IgG antibodies were measured using Human IgA/IgG ELISA Kit (Novus Biologicals, Colorado, United States) according to the manufacturer’s instructions.

Techniques: Clinical Proteomics, Competitive ELISA, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Journal of Virology

Article Title: Infectivity of Hepatitis C Virus Is Influenced by Association with Apolipoprotein E Isoforms

doi: 10.1128/jvi.01063-10

Figure Lengend Snippet: FIG. 4. Suppression of LDLR and SR-BI results in reduced HCV infection. (A) Knockdown of the LDLR and SR-BI by siRNA. HuH7.5 cells were seeded onto 24-well plates and transfected with siRNA (si-control, si-LDLR, and/or si-SR-BI). Twenty-four hours after transfection, cell lysates were analyzed for the expression of the LDLR, SR-BI, and actin by Western blotting. (B) Analysis of HCV infectivity for HuH7.5 cells in which either the LDLR, SR-BI, or both were silenced. Cells were transfected with siRNA (si-control, si-LDLR, and/or si-SR-BI). Twenty-four hours after transfection, HCVcc produced from cells bearing ApoE2 (open bars) or ApoE3 (filled bars) was serially diluted and inoculated. Forty-eight hours after infection, infected cells were counted by fluorescence microscopy after staining with an anti-NS5A antibody. HCV/ApoE3, HCV bearing ApoE3; HCV/ApoE2, HCV bearing ApoE2. P values were determined by comparison (by Student’s t test) with si-control-treated cells. *, P 0.0005; **, P 0.005. (C) HuH7.5 cells were preincubated with control goat IgG (Ctrl-IgG), control rat preimmune serum (Ctrl-serum), or a goat anti-LDLR, rat anti-SR-BI, rat anti-claudin 1 (anti-CLDN1), or mouse anti-CD81 antibody for 1 h at 37°C before infection with serially diluted HCVcc from HuH7.5 cells expressing ApoE2 (open bars) or ApoE3 (filled bars). Forty-eight hours after infection, infected cells were stained with anti-NS5A antibodies and counted by fluorescence microscopy. The average values for three independent experiments are shown; error bars, standard deviations of the means. P values were determined by comparison (by Student’s t test) against cells treated with preimmune serum. *, P 0.0005; **, P 0.005.

Article Snippet: Rat anti-HA (3F10; Roche Applied Science), mouse anti-Flag (M2; Sigma), mouse anti-actin (AC-40; Sigma), goat anti- 1-antitrypsin (K15600G; Biodesign International), mouse anti-ApoE (13F45; Autogen Bioclear), goat anti-ApoE (AB947; Chemicon International), sheep anti-ApoB (K90086C; Biodesign International), mouse anti-core (CP11; Institute of Immunology), goat anti-LDLR (AF2148; R&D Systems), rabbit anti-SR-BI (EP1556Y; Abcam), mouse anti-CD81 (JS-81; BD Biosciences), and normal goat IgG (sc-2028; Santa Cruz Biotechnology) antibodies were purchased commercially.

Techniques: Infection, Knockdown, Transfection, Control, Expressing, Western Blot, Produced, Microscopy, Staining, Comparison

Journal: Immunobiology

Article Title: c-Rel is dispensable for the differentiation and functional maturation of M cells in the follicle-associated epithelium

doi: 10.1016/j.imbio.2016.09.008

Figure Lengend Snippet: Primers used for RT-qPCR analysis.

Article Snippet: For the detection of SpiB in paraformaldehyde-fixed sections, antigen retrieval was performed with citrate buffer (pH 7.0, 121 °C, 5 min) prior to immunostaining with sheep anti-mouse SpiB polyclonal antibody (R&D Systems).

Techniques:

Journal: Immunobiology

Article Title: c-Rel is dispensable for the differentiation and functional maturation of M cells in the follicle-associated epithelium

doi: 10.1016/j.imbio.2016.09.008

Figure Lengend Snippet: SpiB expression in the FAE is unaffected in the absence of c-Rel. (A) IHC analysis of SpiB expression (green) in the FAE of c-Rel −/− and wild-type (WT) control mice. Boxed areas in upper panels are shown at higher magnification in the lower panels. Broken lines indicate the FAE boundaries. Arrows, Spi-B + cell nuclei in the FAE. (B) Morphometric analysis showed that the number of SpiB + cells in the FAE of c-Rel −/− and WT were similar. (C) RT-qPCR analysis suggested there was no significant difference in the expression of Spib mRNA levels in Peyer’s patches from c-Rel −/− or WT mice. Gene expression data are normalised so that the mean level in samples from WT mice was 1.0. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: For the detection of SpiB in paraformaldehyde-fixed sections, antigen retrieval was performed with citrate buffer (pH 7.0, 121 °C, 5 min) prior to immunostaining with sheep anti-mouse SpiB polyclonal antibody (R&D Systems).

Techniques: Expressing, Control, Quantitative RT-PCR, Gene Expression

Journal: The FEBS journal

Article Title: Cadherin-6 controls neuronal migration during mouse neocortical development via an integrin-mediated pathway.

doi: 10.1111/febs.70150

Figure Lengend Snippet: Fig. 1. CDH6 protein is detected in the developing mouse cerebral cortex. (A–G) Immunostaining for CDH6 (green) and DAPI (magenta) using sections from the cerebral cortex at E12.0 (A) (n = 6), E14.0 (B) (n = 6), E16.0 (C) (n = 4) and E18.0 (D–F) (n = 10) of the ICR mouse. The boxed regions in (D) are shown at higher magnification in (E) and (F). (G) Control staining with normal sheep IgG instead of CDH6 antibody (n = 10). The lower panels in (A–G) show grayscale, single-channel images of CDH6 (A–F) and normal sheep IgG (G), respectively. (H–J) The cerebral cortices (E17.0) that had been electroporated with CAG-EGFP plasmid on E14.0 were stained for DCX (H, I) (n = 4) and Nestin (NES) (n = 4) (J). Arrows in (H) and (I) indicate colocalization of immunosignals of CDH6/DCX in EGFP-positive neurons in CP and IZ, respectively. Cell outline is highlighted using the GFP signal as a guide in the CDH6 and DCX panels (magenta lines in H and I). The second panels from the left in (H–J) show CDH6 grayscale, single-channel images. The fourth panels from the left in (H–I) show DCX grayscale, single-channel images. The fourth panel from the left in (J) shows a NES grayscale, single-channel image. (K–M) Immunostaining for HA (green) and DAPI (magenta) using sections from the cerebral cortex at P0 of the Cdh6-HA KI mouse (n = 4). Boxed regions in (K) are shown at higher magnification in (L) and (M). (N) Control staining with normal rat IgG instead of HA antibody (n = 4). The lower panels in (K–N) show grayscale, single-channel images of CDH6 (K–M) and normal rat IgG (N), respectively. Scale bars: 100 lm in (A, B, C, D, G, K, N); 40 lm in (E, F, L, M); 5 lm in (H, I); 20 lm in (J). CP, cortical plate; DCX, Doublecortin; HP, hippocampus; IZ, intermediate zone; MAZ, multipolar cell accumulation zone; MZ, marginal zone; Ncx, neocortex; NES, Nestin; SLM, stratum lacunosum moleculare; SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum; VZ, ventricular zone.

Article Snippet: For control staining for CDH6 and HA, normal sheep IgG (ChromPure sheep IgG, whole molecule; Jackson ImmunoResearch Laboratories, cat# 013-000-003) and normal rat IgG (ChromPure rat IgG, whole molecule; Jackson ImmunoResearch Laboratories, cat# 102-000-003) were used, respectively.

Techniques: Immunostaining, Control, Staining, Plasmid Preparation