Journal: Scientific Reports
Article Title: Neu-medullocytes, sialidase-positive B cells in the thymus, express autoimmune regulator (AIRE)
Figure Lengend Snippet: Antigens expressed in Neu-medullocytes as B cells: IgG, CD5, IgM, and MHC class II. ( I ) Reconfirmation of the expression of IgG on X-NANA-stained cells using F(ab′) 2 fragment of anti-mouse IgG as detected by confocal microscopy. The thymus from a C57BL/6 mouse (male, 6 W) was used. ( A ) X-NANA-stained; ( B ) FITC-anti-mouse IgG; ( C and D ) merged ( A and B ). ( D and E ) enlarged area enclosed by a white square in ( C ) and its DIC (differential interference contrast) image, respectively. ( F ) DIC image including the areas from ( A – C ) (white square) and ( D , E ) (black square). Scale bars in ( A , D and F ) indicate 50 (for A – C ), 20 (for D and E ) and 100 µm (for F ), respectively. ( II ) Other antigens expressed on X-NANA stained cells. The thymus sections from C57BL/6 mice were used for ( A – C ) (male, 6 W). The thymus sections from AKR mice (haplotype k) were used for ( D – F ) (male, 8 W) and for ( G – I ) (female, 5 W). ( A , D and G ) X-NANA stained; ( B ) FITC-anti-CD5 stained; ( C ) merged ( A and B) ; ( E ) FITC-anti-MHC class II (anti I-A k ) stained; ( F ) merged ( D and E ); ( H ) R.R.-anti-IgM stained; ( I ) merged ( G and H ). The scale bars (μm) indicate 20 in A (for A – C ), 20 in D (for D – F ), and 50 in G (for G – I ).
Article Snippet: The following antibodies were purchased: FITC-conjugated F(ab′)2 fragment of donkey anti-mouse IgG (H + L) (Jackson ImmunoResearch, West Grove, PA), anti-CD5 (Q-20, goat polyclonal IgG for mouse CD5 at the N-terminus, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), anti-NEU1 middle region antibody (rabbit polyclonal antibody, Aviva Systems Biology (San Diego, CA)), Anti-AIRE (rabbit polyclonal IgG against AIRE (KLH-conjugated synthetic peptide derived from human AIRE, purified by protein A (cross-reactive species: human, mouse, rat), from Bioss (Boston, MA)).
Techniques: Expressing, Staining, Confocal Microscopy, Mouse Assay