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Figure 4. Inhibition of UFMylation have a role in the prevention of cardiovascular system aging (A–E) 24 (aged) months mice were administered PBS or compound 8.5 (0.25 mg/kg/d) intraperitoneally for 3 weeks. p values correspond to unpaired two-tailed Student’s t test. (A) Quantitative analysis of strain and pulse wave velocity (PWV) in the left carotid artery (LCA), aortic arch, and abdominal aorta of mice, n = 8. (B) Masson and HE staining of aortic cross section. (C) Quantitative analysis of Masson staining, n = 6. (D and E) Immunoblotting and quantification of the indicated protein from mouse aortic endothelial cells, n = 6. All data are represented as mean ± SEM. **, p % 0.01; ***, p % 0.001; ****, p % 0.0001.
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Figure 4. Inhibition of UFMylation have a role in the prevention of cardiovascular system aging (A–E) 24 (aged) months mice were administered PBS or compound 8.5 (0.25 mg/kg/d) intraperitoneally for 3 weeks. p values correspond to unpaired two-tailed Student’s t test. (A) Quantitative analysis of strain and pulse wave velocity (PWV) in the left carotid artery (LCA), aortic arch, and abdominal aorta of mice, n = 8. (B) Masson and HE staining of aortic cross section. (C) Quantitative analysis of Masson staining, n = 6. (D and E) Immunoblotting and quantification of the indicated protein from mouse aortic endothelial cells, n = 6. All data are represented as mean ± SEM. **, p % 0.01; ***, p % 0.001; ****, p % 0.0001.
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Figure 4. Inhibition of UFMylation have a role in the prevention of cardiovascular system aging (A–E) 24 (aged) months mice were administered PBS or compound 8.5 (0.25 mg/kg/d) intraperitoneally for 3 weeks. p values correspond to unpaired two-tailed Student’s t test. (A) Quantitative analysis of strain and pulse wave velocity (PWV) in the left carotid artery (LCA), aortic arch, and abdominal aorta of mice, n = 8. (B) Masson and HE staining of aortic cross section. (C) Quantitative analysis of Masson staining, n = 6. (D and E) Immunoblotting and quantification of the indicated protein from mouse aortic endothelial cells, n = 6. All data are represented as mean ± SEM. **, p % 0.01; ***, p % 0.001; ****, p % 0.0001.
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Figure 4. Inhibition of UFMylation have a role in the prevention of cardiovascular system aging (A–E) 24 (aged) months mice were administered PBS or compound 8.5 (0.25 mg/kg/d) intraperitoneally for 3 weeks. p values correspond to unpaired two-tailed Student’s t test. (A) Quantitative analysis of strain and pulse wave velocity (PWV) in the left carotid artery (LCA), aortic arch, and abdominal aorta of mice, n = 8. (B) Masson and HE staining of aortic cross section. (C) Quantitative analysis of Masson staining, n = 6. (D and E) Immunoblotting and quantification of the indicated protein from mouse aortic endothelial cells, n = 6. All data are represented as mean ± SEM. **, p % 0.01; ***, p % 0.001; ****, p % 0.0001.
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Figure 4. Inhibition of UFMylation have a role in the prevention of cardiovascular system aging (A–E) 24 (aged) months mice were administered PBS or compound 8.5 (0.25 mg/kg/d) intraperitoneally for 3 weeks. p values correspond to unpaired two-tailed Student’s t test. (A) Quantitative analysis of strain and pulse wave velocity (PWV) in the left carotid artery (LCA), aortic arch, and abdominal aorta of mice, n = 8. (B) Masson and HE staining of aortic cross section. (C) Quantitative analysis of Masson staining, n = 6. (D and E) Immunoblotting and quantification of the indicated protein from mouse aortic endothelial cells, n = 6. All data are represented as mean ± SEM. **, p % 0.01; ***, p % 0.001; ****, p % 0.0001.

Journal: iScience

Article Title: UFMylation maintains YAP stability to promote vascular endothelial cell senescence.

doi: 10.1016/j.isci.2025.111854

Figure Lengend Snippet: Figure 4. Inhibition of UFMylation have a role in the prevention of cardiovascular system aging (A–E) 24 (aged) months mice were administered PBS or compound 8.5 (0.25 mg/kg/d) intraperitoneally for 3 weeks. p values correspond to unpaired two-tailed Student’s t test. (A) Quantitative analysis of strain and pulse wave velocity (PWV) in the left carotid artery (LCA), aortic arch, and abdominal aorta of mice, n = 8. (B) Masson and HE staining of aortic cross section. (C) Quantitative analysis of Masson staining, n = 6. (D and E) Immunoblotting and quantification of the indicated protein from mouse aortic endothelial cells, n = 6. All data are represented as mean ± SEM. **, p % 0.01; ***, p % 0.001; ****, p % 0.0001.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-UFM1 Abcam Cat# ab109305; RRID:AB_10864675 Rabbit monoclonal anti-YAP Cell Signaling Technology Cat# 14074; RRID:AB_2650491 Mouse monoclonal anti-YAP Santa Cruz Biotechnology Cat# sc-101199; RRID:AB_1131430 Rabbit monoclonal anti-P53 Cell Signaling Technology Cat# 2527; RRID:AB_10695803) Mouse monoclonal anti-Alpha Tubulin Proteintech Cat# 66031-1-Ig; RRID:AB_11042766 Rabbit monoclonal anti-p16 INK4A Cell Signaling Technology Cat# 18769; RRID:AB_2935679 Rabbit monoclonal anti-DYKDDDDK Tag Cell Signaling Technology Cat# 14793; RRID:AB_2572291 Rabbit isotype control Cell Signaling Technology Cat# 2729; RRID:AB_1031062 Rabbit polyclonal anti-p21 Proteintech Cat# 10355-1-AP; RRID:AB_2077682 Rabbit polyclonal anti-UBA5 ABclonal Cat# A15514; RRID:AB_2762915 Rabbit monoclonal anti-VCAM1 Huabio Cat# ET1601-18; RRID:AB_3069600 Rabbit polyclonal anti-UFL1 Oasis Biofarm Cat# OB-PRB036; RRID:AB_2938995 Anti-rabbit IgG, HRP-linked Antibody Cell Signaling Technology Cat# 7074; RRID:AB_2099233 Anti-mouse IgG, HRP-linked Antibody Cell Signaling Technology Cat# 7076; RRID:AB_330924 Bacterial and virus strains MCS-3xFLAG-EGFP-YAP adenovirus GENECHEM N/A Chemicals, peptides, and recombinant proteins Cycloheximide MedChemExpress Cat#HY-12320 Verteporfin MedChemExpress CatHY-B0146 Compound 8.5 MedChemExpress Cat#HY-148266 ECM culture medium Zhong Qiao Xin Zhou Biotechnology Cat#ZQ-1304 Trypsin-EDTA Thermo Fisher Scientific Cat#25200056 Protease inhibitor cocktail tablets Roche Cat#04693132001 Phosphatase inhibitor cocktail tablets Roche Cat#04906845001 PMSF Solarbio Life Sciences Cat#IP0280 Critical commercial assays TransZol Up Plus RNA Kit TransGenBiotech Cat# ER501-01 Lipofectamine RNAiMAX Transfection Reagent Invitrogen Cat# 13778030 Pierce BCA Protein Assay Kit Thermo Fisher Scientific Cat# 23225 ECL Western Blotting Detection Kit Affinity Cat# KF003 Senescence b-Galactosidase Staining Kit Beyotime Biotechnology Cat# C0602 Lipofectamine 3000 Transfection Reagent Thermo Fisher Scientific Cat# L3000001 Deposited data RNA-seq This paper GEO: GSE273828 Source data This paper https://data.mendeley.com/preview/ 2c97jkvx9c?a=f6996a71-5f0f-469d84af-f85dbb1ea464 Experimental models: Cell lines HEK293T ATCC CRL-1573; RRID: CVCL0045 HUVEC (pooled donor) endothelial cells Lonza Cat# CC-2519 Experimental models: Organisms/strains Mouse C57BL6J Jackson Laboratories N/A (Continued on next page) iScience 28, 111854, February 21, 2025 e1

Techniques: Inhibition, Two Tailed Test, Staining, Western Blot