Figure 1.

Binding of serum antibodies to mouse fibroblasts indicates presence of anti-fibroblast antibodies in serum from asbestos-instilled mice. (A) Mouse primary skin fibroblasts were fixed with paraformaldehyde and stained using serum from (left) saline- or (right) tremolite-exposed mice as the primary antibody, followed by incubation with Alexa-488-conjugated anti-mouse IgG secondary Ab. Nuclei are stained with propidium iodide. Images (at 400×) are representative of multiple experiments. (B) A cell-based ELISA was performed using L929 mouse fibroblasts, to measure the binding of serum antibodies to the cells. As in A, binding of serum antibodies was compared between sera from saline- or tremolite-treated mice. N = 7, *P < 0.05 by unpaired 2-tailed t-test. (C) The cell-based ELISA was repeated using L929 cells, and including a positive control anti-fibroblast antibody (anti-Fb, anti-prolyl-4-hydroxy-lase), as well as serum from tremolite-exposed mice, either uncleared of IgG (trem unclr) or that had been cleared of IgG by Protein G precipitation (cleared). N = 4, *P < 0.05 compared to secondary antibody-only control. (See colour version of this figure online at www.informahealthcare.com/imt)