Journal:

Article Title: Enteric Expression of the Integrin ? v ? 6 Is Essential for Nematode-Induced Mucosal Mast Cell Hyperplasia and Expression of the Granule Chymase, Mouse Mast Cell Protease-1

doi:

Figure Lengend Snippet: a: H&E-stained exfoliated jejunal epithelium from S129 mice, showing the relative absence of contaminating cells from the lamina propria (horizontal bar, 20 μm). b and c: Sections of jejunum from an S129 mouse taken on day 7 after infection with N. brasiliensis, stained with rabbit anti-integrin β6 IgG B1, specific for integrin β6 (b) or normal rabbit control IgG (c) (horizontal bar, 10 μm). Note the predominantly circumferential immunofluorescence in the crypt epithelium. Confocal images were acquired in fast photon-counting mode (300 accumulated frames) and have not been subjected to contrast enhancement of any form. d: RT-PCR products for TGF-β1 (405 bp), integrin β6 (340 bp), IgA (316 bp), and the housekeeping gene GAPDH (430 bp) from total RNA extracted from whole jejunum on day 7 after challenge with N. brasiliensis (WJ), and from isolated epithelium taken on day 0 (0-1 to 0-3), day 5 (5-1 to 5-3), day 7 (7-1 to 7-3), and day 9 (9-1 to 9-3) after infection with N. brasiliensis and a negative control with nonreverse-transcribed RNA only (c). Note that transcripts for β6 are apparently less abundant in whole jejunum than in isolated epithelium and that, conversely, the intensity of RT-PCR signals for TGF-β1 and for IgA are stronger in samples of whole jejunum (WJ) with little or no signal for IgA in isolated epithelium. e: Relative abundance of mast cell protease transcripts in isolated epithelium from S129 mice. The graphs show the intensity of each PCR product as a proportion of corresponding GAPDH product from the same sample. Data are shown from samples taken on day 0 (uninfected) (open columns), day 5 (horizontally lined columns), day 7 (hexagonally lined columns), and day 9 (filled columns) after infection with N. brasiliensis. Primers were for mMCP-1 (M1) and mMCP-2 (M2). (dOvsd9: ∗p < 0.05, ∗∗p < 0.01).

Article Snippet: Sections were then incubated overnight at 4°C in a humidified container with rabbit anti-integrin β 6 IgG (B1) tissue culture supernatant 13 or 0.5 μg/ml of normal rabbit control IgG (Cambridge Bioscience, Cambridge, UK) in high-salt/10% normal donkey serum.

Techniques: Staining, Infection, Control, Immunofluorescence, Reverse Transcription Polymerase Chain Reaction, Isolation, Negative Control