File S1 - Calpain-2 Compensation Promotes Angiotensin II-Induced Ascending and Abdominal Aortic Aneurysms in Calpain-1 Deficient Mice

File includes Figures S1–S6. Figure S1. Genotyping of experimental mice for calpain-1 alleles by PCR. Genomic DNA from tail biopsies was isolated and screened by PCR for calpain-1 wild type (+/+) and null (−/−) alleles. Reaction products were sized using agarose gel electrophoresis. Figure S2. Calpain-1 deficiency did not affect lipoprotein cholesterol distributions. Lipoproteins were resolved by size-exclusion chromatography. Total cholesterol concentrations are expressed as mean absorbance per fraction. Symbols represent the means and bars are SEMs of 5 individual mice per group: Calpain-1 +/+ (circles) and calpain-1 −/− (triangles). Figure S3. Examples of vascular pathology measurements. Ultrasound images (A - Day 0 and Day 28), ex vivo pictures of suprarenal aortas (B - after termination) and ascending arch aortas (C), that represent aortic diameters nearest the mean of each group. Figure S4. Calpain-1 deficiency had no effect on AngII-induced atherosclerosis in LDL receptor −/− mice. Atherosclerotic lesion areas were measured on aortic arch intimal surfaces (n = 18–19). Open circles (calpain-1 +/+) and gray circles (calpain-1 −/−) represent individual mice, diamonds represent means, and bars are SEMs. Statistical analyses were performed with nonparametric Mann-Whitney Rank sum test. Figure S5. Histological and cellular characteristics of AngII-induced abdominal AAs in calpain-1 +/+ and calpain-1 −/− mice. Representative suprarenal aortic tissue-sections from AngII infused calpain-1 +/+ and calpain-1 −/− mice stained with Movat’s pentachrome (A–D) and immunostained for CD68 (E–H). Elastin stains black; CD68+ cells stain red. Scale bars corresponds to 50 µm. Arrow indicates medial break and positive staining with CD68. A,C,E and G = 40×; B,D,F and H = 200×. Figure S6. Histological and cellular characteristics of AngII-induced ascending AAs in calpain-1 +/+ and calpain-1 −/− mice. Representative anterior ascending aortic tissue-sections from AngII infused calpain-1 +/+ and calpain-1 −/− mice stained with Movat’s pentachrome (A–D) and immunostained for CD68 (E–H), Elastin stains black; CD68+ cells stain red. Scale bars corresponds to 50 µm. Arrow indicates medial break and positive staining with CD68. A,C,E and G = 40×; B,D,F and H = 200×. (PDF)