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Blood, Vol. 111, Issue 9, 4596-4604, May 1, 2008
The CD40-TRAF6 axis is the key regulator of the CD40/CD40L system in neointima formation and arterial remodeling
Blood Donners et al.
111: 4596
Supplemental materials for: Donners et al
Files in this Data Supplement:
- Figure S1. Knockout and transgenic mice with defects in CD40 signaling (JPG, 90.3 KB)
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(A) Besides wt, CD40−/−, CD40L−/− we used CD40−/− mice that express a human CD40 construct under a MHC-II promoter. Mutations in TRAF-binding domains of CD40 disrupt binding of specific TRAFs to CD40. wt: wild-type C57BL6/J mice, TRAF-wt: CD40−/− mice expressing normal human CD40 under the MHC-II promoter, CD40-T2/3/5: mice with a defect in binding of TRAF2,3 and 5 to CD40, CD40-T6: mice with a defect in binding of TRAF6 to CD40, CD40-T2/3/5&6: mice with a defect in both binding domains for TRAF2/3/5 and TRAF6. (B) Immunohistochemical staining showed MHC-II expression in neointimal lesions.
- Figure S2. Baseline geometry of the contralateral, non-ligated artery does not differ between the genotypes (JPG, 105 KB)
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- Figure S3. Baseline geometry and SMC content of a normal right carotid artery does not differ between the genotypes (JPG, 90.1 KB)
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- Figure S4. Expression of TRAF2 (A), TRAF3 (B) and TRAF6 (C) in neointimal lesions (JPG, 251 KB)
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All TRAFs are predominantly expressed in the neointima, but also in the media (M).
- Figure S5. Panel showing double IHC with MMP-2 (blue) or MMP-9 with Mac3 (red) in neo-intimas of wild type (WT) and CD40−/− mice (JPG, 397 KB)
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Macrophages in neo-intimas of both genotypes express MMP-2 and -9. In the CD40−/− group, less macrophages are present, resulting in decreased MMP-2 and -9 expression.
- Figure S6. Leukocyte-endothelium adhesion assay, revealing that leukocytes deficient in CD40 have an impaired adhesion capability to TNFα stimulated endothelial cells (SVEC) compared to wild type leukocytes (JPG, 61.6 KB)
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