Journal: Immunology

Article Title: Lack of Shiga-like toxin binding sites in germinal centres of mouse lymphoid tissues

doi: 10.1046/j.1365-2567.2002.01373.x

Figure Lengend Snippet: Lack of Shiga-like toxin (SLT)-binding sites in germinal centres of the lymph nodes and Peyer’s patches from ICR mice. Acetone-fixed frozen sections of normal mouse kidneys [(a), (b) and (g)], mouse draining lymph nodes 6 days after subcutaneous immunization [(c), (d) and (e)], or mouse Peyer’s patches 4 days after oral immunization [(f) and (h),], were stained with digoxigenin-labelled SLT-1B proteins (DIG-SLT-1B) plus horseradish peroxidase (HRP)-anti-DIG [(a) and (b)]. The sections were then counterstained with haematoxylin. The other sections were stained with haematoxylin and eosin (c); biotin-conjugated peanut agglutinin (PNA) plus Texas Red avidin D (d) and (f); or DIG-SLT-1B plus fluorescein isothiocyanate (FITC)-anti-DIG (e), (g) and (h).(a): Cortical regions of the kidney. Renal tubules were strongly stained with DIG-SLT-1B (arrowheads) while glomeruli were not stained (single arrow).(b): Medullary regions of the kidney. Cross-sections of collecting ducts were strongly stained with DIG-SLT-1B.(d), (e), (f) and (h): Germinal centres (arrows) were strongly stained with PNA but not with DIG-SLT-1B. Arrowheads point towards the intestinal epithelium (f) and (h).(g): Collecting ducts in kidney medulla were strongly stained with DIG-SLT-1B, as revealed by immunofluorescence. Bars represent 50 µm [(a) and (b), × 230 magnification] or 200 µm [(c) to (h), × 70 magnification].

Article Snippet: Texas Red-avidin D, HRP-avidin D, biotin-conjugated peanut agglutinin (PNA) and 3-amino-9-ethylcarbazole (AEC) substrate kit were obtained from Vector (Burlingame, CA).

Techniques: Binding Assay, Staining, Avidin-Biotin Assay, Immunofluorescence

Journal: Immunology

Article Title: Lack of Shiga-like toxin binding sites in germinal centres of mouse lymphoid tissues

doi: 10.1046/j.1365-2567.2002.01373.x

Figure Lengend Snippet: Lack of Shiga-like toxin (SLT)-binding sites on peanut agglutinin (PNA)-positive B cells from immunized ICR mouse lymph nodes. Draining lymph node cells obtained 6 days after subcutaneous immunization were stained with phycoerythrin (PE)-anti-B220 (FL-2) and biotin-PNA/PC5-streptavidin (FL-4). Cells were also treated with (a)and (b)or without(c) and (d)digoxigenin-labelled SLT-1B proteins (DIG-SLT-1B). The binding of DIG-SLT-1B was detected by fluorescein isothiocyanate (FITC)-anti-DIG (FL-1). The gated populations representing PNAbright/B220+ cells (the square in panel a) were analysed for the binding of DIG-SLT-1B (b). FL-1 signals from PNAbright/B220+-gated cells, without treatment of DIG-SLT-1B (the square in panel c), were shown as a background (d). The binding sites for SLT on RAMOS cells were demonstrated by treatment with (f) or without (e) DIG-SLT-1B, followed by incubation with FITC-anti-DIG (FL-1).

Article Snippet: Texas Red-avidin D, HRP-avidin D, biotin-conjugated peanut agglutinin (PNA) and 3-amino-9-ethylcarbazole (AEC) substrate kit were obtained from Vector (Burlingame, CA).

Techniques: Binding Assay, Staining, Incubation

Journal: Scientific Reports

Article Title: Central expression of synaptophysin and synaptoporin in nociceptive afferent subtypes in the dorsal horn

doi: 10.1038/s41598-019-40967-y

Figure Lengend Snippet: Co-localization of synaptophysin and synaptoporin with IB4-binding C fibers in the thoracic dorsal horn. IB4 binding C fibers distributed in lamina II of T7 and T13 dorsal horns and expressed synaptophysin and/or synaptoporin as shown in a representative montage of confocal images from T7. ( A ) Areas of IB4 binding projections within the investigated region of interest were analyzed at both thoracic levels.( B ) Colocalization of IB4 binding axon terminals with synaptophysin or with synaptoporin was assessed by double labeling quantification ( C ) to find significantly different expression levels of synaptoporin and synaptophysin in this C fiber subpopulation (n = 4, p < 0.05, Kruskal-Wallis test with multiple comparison test, Cohen’s d > 1.3).

Article Snippet: The primary antibodies included goat anti-IB4 (1:400, Vector, Burlingame, CA), goat anti-CTB (1:2000, List Biological Laboratories, Campbell, CA), mouse anti-synaptophysin (1:1000, Synaptic Systems), rabbit anti-synaptoporin (1:300, Synaptic Systems), and Guinea pig anti-CGRP (1:2000, Peninsula Laboratories International, San Carlos, CA).

Techniques: Binding Assay, Labeling, Expressing

Journal: Scientific Reports

Article Title: Central expression of synaptophysin and synaptoporin in nociceptive afferent subtypes in the dorsal horn

doi: 10.1038/s41598-019-40967-y

Figure Lengend Snippet: Expression of synaptophysin and synaptoporin in dorsal cutaneous C fibers in the T7 and T13 spinal cord. Cutaneous C fibers were labeled with IB4 injections into DCNs. ( A ) DCN-specific C fibers projected laterally at the inner layer of laminae II overlaid by the synaptoporin distribution area as shown in representative montages of confocal images at T7. ( B ) Overlapping areas of DCN-specific IB4+ C fibers with synaptoporin and synaptophysin were quantified at every 200 μm across the sections centered at the dorsal horn entry zone (DREZ) at T7 and T13. ( C ) Bar graphs show averaged percent overlapping areas across those serial sections from 3 rats ( D ).

Article Snippet: The primary antibodies included goat anti-IB4 (1:400, Vector, Burlingame, CA), goat anti-CTB (1:2000, List Biological Laboratories, Campbell, CA), mouse anti-synaptophysin (1:1000, Synaptic Systems), rabbit anti-synaptoporin (1:300, Synaptic Systems), and Guinea pig anti-CGRP (1:2000, Peninsula Laboratories International, San Carlos, CA).

Techniques: Expressing, Labeling