Fig. 2. Disruption of Zbtb7a expression in lungs prevents anti-MHC-induced lung-restricted autoimmunity and OAD.

(A) Schematic representation of si-RNA lentivirus transduction and subsequent Ab challenges. Mice received an intrabronchial delivery of lentivirus (10⁷ IU/mouse) three days prior to intrabronchial Ab regimen. (B) Repression in the Zbtb7a expression was quantified (as described in Fig. 1C). (C) Detection of Zbtb7a expression by Western blot analysis in lung lysates of three mice by anti-Zbtb7a (clone 466407) and anti-β-Actin (clone C4). (D) Representative Masson’s trichrome staining on day 45 post-Ab administration. Fibrotic areas with deposition of collagenous ECM are stained blue, (b, bronchiole; o, occluded bronchiole). (E) Histopathologic score for each data point represents average of five fields from each mouse (19). Median lines are drawn. (F) Titers for serum anti-Kα1T and -Col V were evaluated by ELISA from eight mice and mean±SEM lines are drawn. (G) Development of T cell autoreactivity was analyzed by ELISPOT on day 45. The data are presented as spot-forming cell (SFC) per million cells. Results are presented as mean±SEM. Multiple t-tests were applied with correction for Holm-Sidak method and significance of difference is marked (p values indicated). Scale bar in panel D is 100µm.