Journal: Disease Models & Mechanisms

Article Title: Alterations in nuclear structure promote lupus autoimmunity in a mouse model

doi: 10.1242/dmm.024851

Figure Lengend Snippet: Anti-neutrophil serum reactivity develops in (NZW×B6.Lbr ic )F 1 mice. (A) Fluorescence microscopy of ethanol-fixed human PMNs stained with DAPI (top panel), and female (NZW×B6.Lbr ic )F 1 mouse serum and an Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (bottom panel). Broken circles indicate DAPI-positive bilobed eosinophils that were not readily detected by the female (NZW×B6.Lbr ic )F 1 mouse serum. Image magnification, 100×. (B) High-power magnification of ethanol-fixed PMN staining. Neutrophils displayed strong nuclear staining, but eosinophils had comparatively weak and patchy perinuclear staining. Image magnification, 400×. The preferential staining of isolated neutrophils relative to peripheral blood mononuclear cells and isolated monocytes is demonstrated in greater detail in Fig. S7 . (C) Ethanol-fixed PMNs were co-stained with female (NZW×B6.Lbr ic )F 1 mouse serum (green) and anti-MPO (red), nuclei were counterstained with DAPI (blue). The mouse serum staining did not colocalize with MPO. Image magnification, 1000×.

Article Snippet: HEp-2 cells immobilized on microscopy slides (MBL/Bion) were incubated with a 1:200 dilution of mouse sera, and a FITC-conjugated goat anti-mouse immunoglobulin secondary antibody (1:1000; 1010-02, Southern Biotech).

Techniques: Fluorescence, Microscopy, Staining, Isolation