Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: Immunocytochemical detection of MMCs in epithelial whole mounts and frozen sections of jejunum from T. spiralis-infected BALB/c mice. A to D: Whole mounts of jejunal epithelium were probed with anti-mMCP-1 (pseudo-colored red) or isotype-matched control antibodies together with anti-IgE (pseudo-colored green) or isotype-matched control antibodies and counterstained with DRAQ5 (pseudo-colored blue). IgE surface labeling was almost exclusively restricted to mast cells exhibiting strong intracellular mMCP-1 staining in samples probed with mMCP-1- and IgE-specific antibodies (A and D). Single-negative controls for IgE (B) and mMCP-1 (C) were used to confirm labeling specificity. Frozen sections of jejunum from uninfected (E) and T. spiralis-infected (F) BALB/c mice were probed with rabbit anti-tryptase (pseudo-colored green), anti-IgE (pseudo-colored red), and anti-integrin-α6 antibodies (pseudo-colored blue). No IgE or tryptase-positive cells were detected in uninfected animals (E). Abundant IgE+ve cells were detected in the samples from infected mice, the majority of which were located intraepithelially (F). Although significant numbers of intraepithelial IgE+ve cells were tryptase-negative (F, arrowheads), most IgE+ve cells detected in the lamina propria expressed tryptase (F, arrow). Staining specificity was confirmed using single-positive controls for tryptase (G), integrin-α6 (H), and IgE (I), in which the two other antigen-specific antibodies were replaced with control IgG. Identical excitation and exposure settings were used to acquire images of multilabeled and control samples. Scale bar length (μm) is provided in the bottom left corner of each image.

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Infection, Labeling, Staining

Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: IgE+ve cells in the jejunal mucosa of T. spiralis-infected mice express integrin-αEβ7. Frozen sections of jejunum from infected BALB/c mice were triple-labeled with anti-integrin-αE or anti-integrin-β7 together with anti-IgE and anti-integrin-α6 (see Figure 4). Using integrin-α6 staining to differentiate the epithelium (filled bars) and lamina propria (open bars), IgE+ve cells mm−2 (A) and the relative frequencies of IgE+ve cells expressing integrin-αE or integrin-β7 (B) were determined for each location using five randomly selected 0.31-mm2 fields of view per sample. The mean percentage of intraepithelial IgE+ve cells ± 1 SEM is provided in parentheses (A). Data are expressed as mean values ± 1 SEM (n = 4). Differences observed between the epithelial and lamina propria compartments were significant at P < 0.01 (**).

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Infection, Labeling, Staining, Expressing

Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: Immunofluorescent detection of integrin-αE and -β7 expression by IgE+ve cells and PCNA-specific staining of toluidine blue-positive mast cells in the jejunal mucosa of S129 β6−/− and β6+/+ mice. A to J: Frozen sections of jejunum were probed with monoclonal rat antibodies specific for integrin-α6 (pseudo-colored blue), IgE (pseudo-colored green), and integrin-β7 or -αE (pseudo-colored red). Representative images are shown from uninfected β6+/+ (A) and β6−/− (B) mice stained for IgE, integrin-α6, and integrin-β7; uninfected β6+/+ (C) and β6−/− (D) mice stained for IgE, integrin-α6, and integrin-αE; T. spiralis-infected β6+/+ (E) and β6−/− (F) mice stained for IgE, integrin-α6, and integrin-β7; and T. spiralis-infected β6+/+ (G) and β6−/− (H) mice stained for IgE, integrin-α6, and integrin-αE. Arrowheads in D and H highlight occasional cells that exhibit low-level integrin-αE immunoreactivity in sections from β6−/− mice (inset in D has been contrast enhanced). Higher magnification images from T. spiralis-infected β6+/+ (I) and β6−/− (J) mice stained for IgE, integrin-α6, and integrin-αE are annotated with arrows (intraepithelial IgE+ve cells) and arrowheads (lamina propria IgE+ve cells). Note that although large numbers of IgE+ve cells are present in the mucosa of T. spiralis-infected β6−/− mice, they are predominantly located in the lamina propria. Single-positive controls from β6+/+ mice for integrin-β7 (K), integrin-αE (L), IgE (M), and integrin-α6 (N), in which two of the three antigen-specific antibodies were substituted with isotype-matched controls, were used to confirm staining specificity. Identical excitation and exposure settings were used to acquire images A to D. Constant excitation and exposure settings were also maintained during acquisition of images E to N. Bright-field images of toluidine blue-labeled mast cells probed with anti-PCNA antibodies from T. spiralis-infected β6+/+ (O) and β6−/− (P) mice, arrow indicates a PCNA-positive mast cell in O. Scale bars, 25 μm.

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Expressing, Staining, Infection, Labeling

Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: IgE+ve cell recruitment and integrin-αEβ7 expression in the jejunal mucosa of T. spiralis-infected β6−/− and β6+/+ S129 mice. Frozen sections of jejunum from infected β6−/− (open bars) and β6+/+ (filled bars) S129 mice were triple-labeled with anti-integrin-αE or anti-integrin-β7 together with anti-IgE and anti-integrin-α6 (see Figure 4). Data are presented as mean values ± 1 SEM (n = 3) for IgE+ve cells mm−2 (A); the percentage of IgE+ve cells located intraepithelially (B); and the relative frequencies of IgE+ve cells expressing integrin-αE (C) or integrin-β7 (D). Differences observed between β6−/− and β6+/+ S129 mice were significant at P < 0.05 (*) and P < 0.01 (**).

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Expressing, Infection, Labeling

Journal:

Article Title: Expression of Integrin-? E by Mucosal Mast Cells in the Intestinal Epithelium and Its Absence in Nematode-Infected Mice Lacking the Transforming Growth Factor-? 1 -Activating Integrin ? v ? 6

doi:

Figure Lengend Snippet: RT-PCR analysis of integrin-αE and -β7 transcript abundance and toluidine blue analysis of MMC distribution in the jejunal mucosa of T. spiralis-infected β6+/+ and β6−/− S129 mice. A: RT-PCR products for integrin-αE, integrin-β7, and the housekeeping gene GAPDH from reverse-transcribed total jejunal RNA from infected β6−/− (lanes 1 to 4) and β6+/+ (lanes 5 to 8) mice. B: RT-PCR product signal intensity expressed as a percentage of GAPDH signal intensity. Differences in PCR product abundance between β6+/+ and β6−/− S129 mice were significant at P < 0.05 (*). Mast cells were identified using toluidine blue staining (pH 0.5) in Carnoy’s-fixed/paraffin-embedded sections from β6+/+ (C) and β6−/− (D) S129 mice. Note that mast cells are present in greater numbers in β6−/− mice but are predominantly restricted to the lamina propria. Scale bars, 50 μm.

Article Snippet: Purified monoclonal rat IgG 2a anti-human integrin-α 6 (clone GoH3), control rat IgG 2a (clone R35-95), control rat IgG 1 (clone R3-34), and FITC-conjugated control rat IgG 1 (clone R3-34) were purchased from BD Biosciences, Cowley, UK.

Techniques: Reverse Transcription Polymerase Chain Reaction, Infection, Staining

Journal: Frontiers in Neuroscience

Article Title: Altered Light Sensitivity of Circadian Clock in Shank3 +/– Mouse

doi: 10.3389/fnins.2021.604165

Figure Lengend Snippet: Immunostaining for cholera toxin β-subunit (CTB) and NMDAR2A in the suprachiasmatic nucleus (SCN) from wild-type (WT) and Shank3 +/– mice. CTB (diaminobenzidine, DAB, left column ) and NMDAR2A ( right column ) did not show obvious differences in the immunoreactivity between groups (WT, top micrographs ; Shank3 +/– , bottom micrographs ). For this and subsequent figures ( , ), scale bar = 50 μm.

Article Snippet: Primary antibodies were diluted as indicated in 0.1 M PBS containing 1.0% normal serum in 0.3% Triton X-100 [anti-VIP raised in rabbit, CAT 20077, Incstar, 1:2,000 ( ); anti-CTB subunit raised in goat, CAT 703, List Biological Laboratories, 1:2,000 ( ); anti-NMDAR2A (GluN2A) raised in rabbit, CAT ACG 002, Alomone Labs, 1:400 ( ); and anti c-FOS raised in rabbit, CAT SC-52, Santa Cruz, 1:1,000 ( )].

Techniques: Immunostaining