Additional file 2 of Cysteine-rich protein 2 deficiency attenuates angiotensin II-induced abdominal aortic aneurysm formation in mice
Creators: name: Chen, Chung-Huang
name: Ho, Hua-Hui
name: Jiang, Wei-Cheng
name: Ao-Ieong, Wai-Sam
name: Wang, Jane
name: Orekhov, Alexander N.
name: Sobenin, Igor A.
name: Layne, Matthew D.
name: Yet, Shaw-Fang
Publisher: name: figshare
Year: 2022
Subjects: subject: Biophysics
subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: Science Policy
subject: 69999 Biological Sciences not elsewhere classified
subject: Developmental Biology
subject: Computational Biology

description: Additional file 2: Fig. S1. Total cholesterol level and aortic morphology in Apoe−/− and Csrp2−/−Apoe–/– mice. Apoe−/− and Csrp2−/−Apoe−/− mice were infused with saline or Ang II and fed a high-fat diet for 4 weeks. A Representative aortas from Apoe−/− (n = 12) and Csrp2−/−Apoe−/− (n = 13) mice infused with saline. B Total cholesterol levels from plasma of Apoe−/− and Csrp2−/−Apoe−/− mice infused with saline (n = 6 and 10, respectively) or Ang II (n = 6 and 5, respectively). C Histological analysis of abdominal aortas from Apoe−/− and Csrp2−/−Apoe−/− mice infused with saline. Aortic sections were stained with H&amp;E, SM α-actin for smooth muscle cells, and Verhoeff’s stain elastin to delineate elastin layers. Fig. S2. Baseline ROS level and MMP activity in the aortic wall of Apoe−/− and Csrp2−/−Apoe−/− mice. Mice were infused with saline for 2 weeks and abdominal aortas harvested for analysis. A DHE staining (red fluorescence) was performed to assess ROS level on abdominal aortic sections. B In situ zymography was performed to measure MMP activity (green fluorescence) on abdominal aortic sections. Med, media; Lu, lumen. Fig. S3. Expression of MMP9, collagen I, and collagen III in the mouse aorta. Apoe−/− and Csrp2−/−Apoe−/− mice were infused with saline for 2 weeks and abdominal aortas harvested for immunohistochemistry. A Abdominal aortic sections were stained with MMP9 antibody. Aneurysmal aortic sections were used as positive control (right panel). Arrows indicate positive staining of MMP9 in the infiltrated immune cells. B Abdominal aortic sections were stained with collagen I (Col I) antibody (left two panels). Right panel, total proteins were prepared from Apoe−/− and Csrp2−/−Apoe−/− mouse aortas for Western blot analysis to detect Col I and CRP2 expression, and α-tubulin was used as a loading control. C Abdominal aortic sections were stained with collagen III (Col III) antibody (left two panels). Right panel, total proteins were prepared from Apoe−/− and Csrp2−/−Apoe−/− mouse aortas for Western blot analysis to detect Col III and CRP2 expression, and α-tubulin was used as a loading control. Fig. S4. CRP2 modulates AngII-induced AAA incidence and blood pressure. Mice were infused with saline or AngII for 4 weeks while on chow diet and aortas harvested for examination of aneurysm formation. A Apoe−/− and B Csrp2−/−Apoe−/− mice were infused with Ang II for 4 weeks. White arrow, abdominal aneurysm. C Saline infusion did not induce AAA formation in Apoe−/− (n = 3) or Csrp2−/−Apoe−/− (n = 2) mice. Ang II infusion for 4 weeks induced AAA in 33% of Apoe−/− mice (n = 9) while no AAA was observed in Csrp2−/−Apoe−/− (n = 7) mice. D Systolic blood pressure of Apoe−/− (n = 4) and Csrp2−/−Apoe−/− (n = 4) mice was measured at baseline and at different days (7, 10, 14, and 21 d) following Ang II infusion. *P