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Blood, Vol. 111, Issue 10, 5017-5027, May 15, 2008
The proapoptotic and antimitogenic protein p66SHC acts as a negative regulator of lymphocyte activation and autoimmunity
Blood Finetti et al.
111: 5017
Supplemental materials for: Finetti et al
Files in this Data Supplement:
- Figure S1. Normal thymocyte and B cell development in 12-month-old mice (JPG, 93.3 KB)
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(A) Flow cytometric analysis of thymocytes from 12-month-old control (+/+) or p66Shc−/− (−/−) mice stained with anti-CD3 and anti-CD4/anti-CD8 fluorochrome-conjugated mAb. Left, representative FACS profiles of CD4/CD8 labeled thymocytes. The percentage of cells that falls into the indicated quadrants is indicated. Right, bar graph showing the mean values ±SD of double negative (DN), double positive (DP) or single positive thymocytes for CD4 and CD8 expression. (B) Flow cytometric analysis of splenocytes from 12-month-old control (+/+) or p66Shc−/− (−/−) mice stained with anti-IgM and anti-IgD fluorochrome-conjugated antibodies. Left, representative FACS profiles of IgM/IgD labeled splenocytes. The percentage of cells that falls into the indicated regions is indicated. Right, bar graph showing the mean values ±SD of mature (IgMlow IgDhigh), T1 (IgMlow IgDlow) or T2 (IgMlow IgDhigh) splenocytes. For each sample fluorescence was analyzed on gated cells with forward and side light scatter properties of lymphocytes (n≥8; *p<0.05,***p<0.001).
- Figure S2. Presence of skin reactive autoantibodies in the sera of p66Shc−/− mice (JPG, 65 KB)
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Immunofluorescence analysis of frozen skin sections from control mice using sera from control (+/+) or p66Shc−/− (−/−) mice as primary antibodies and FITC-conjugated goat anti-mouse immunoglobulins as secondary antibodies. The sera from p66Shc−/− mice positive for anti-dsDNA antibodies were used in these experiments. Note the diffuse dermal staining by the serum from p66Shc−/− mice, similar to the staining obtained with the secondary antibody alone on the skin of alopecic p66Shc−/− mice shown in figure 7D. Scale bar 100 µm.
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