Journal: Cellular and Molecular Life Sciences: CMLS
Article Title: Endothelial CHOP as a central mechanism in renovascular hypertension-induced vascular endothelial dysfunction and cardiac fibrosis
doi: 10.1007/s00018-025-05741-6
Figure Lengend Snippet: CHOP downregulation in human cardiac microvascular endothelial cells via CHOP-specific shRNAplasmid. (A) Western blot analysis of phosphorylated endothelial nitric oxide synthase (p-eNOS) was performed on cultured human cardiac microvascular endothelial cells transfected with CHOP-specific shRNA or a scrambled plasmid for 4 h. Following transfection, cells were stimulated with angiotensin II for an additional 4 h, followed by ATP stimulation for 15 min. (B) Western blot analysis of CHOP, inflammatory markers (COX2 and NLRP3), and β-actin in endothelial cells treated with CHOP-specific shRNA or scrambled plasmid for 4 h and subsequently stimulated with angiotensin II for 4 h. These data highlight the role of CHOP in modulating endothelial nitric oxide signaling and inflammatory responses under angiotensin II stimulation. Data are presented as mean ± SEM. Statistical analysis was performed using One-way ANOVA followed by Tukey’s post hoc test. Non-significant (ns): p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 for comparisons between (A) CL vs. CTL + Ang-II + ATP vs. CTL + ATP, shRNA CHOP vs. shRNA CHOP + Ang-II + ATP vs. shRNA CHOP + ATP vs. shRNA scramble vs. shRNA scramble + Ang-II + ATP vs. shRNA scramble + ATP, (B) CL vs. CTL + Ang-II, shRNA CHOP vs. shRNA CHOP + Ang-II vs. shRNA scramble vs. shRNA scramble + Ang-II ( n = 3)
Article Snippet: 0.3 × 10 6 Human Cardiac Microvascular Endothelial Cells (HMVEC-C, Cat No: CC-7030, from Lonza) were seeded in a six-well plate and grown in EGMTM-2MV BulletKitTM Medium complete media (Cat No: CC-3202, from Lonza) for 24 h at 37 °C, 5% CO 2 , and 95% air.
Techniques: Western Blot, Cell Culture, Transfection, shRNA, Plasmid Preparation
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