Journal: Clinical Science (London, England : 1979)

Article Title: The loss of glycoprotein nonmetastatic melanoma protein B (GPNMB) alters endothelial cell permeability, metabolism, and survival during infectious challenge

doi: 10.1042/CS20256682

Figure Lengend Snippet: (A ) Interleukin-8 (IL-8), ( B ) tumor necrosis factor-alpha (TNF-α), ( C ) the tissue inhibitors of metalloproteinases 1 (TIMP1) concentrations, and ( D ) metalloproteinases-2, ( E ) metalloproteinases-3, and ( F ) metalloproteinases-9 activity in the culture medium of human microvascular endothelial cells (HMVECs) treated with scrambled siRNA (SCR) or GPNMB-targeting siRNA (siGPNMB), following stimulation with either vehicle or heat-killed E. coli (HKEC, 3 × 10⁸ cells/ml) for 6 h. Each dot represents an individual sample. data are presented as mean ± SEM. Statistical analysis: Two-way ANOVA, followed by the Tukey post hoc test.

Article Snippet: Human dermal microvascular endothelial cells (HMVEC) were obtained from the American Type Culture Collection (ATCC CRL-4060; Manassas, VA, U.S.A.).

Techniques: Activity Assay

Journal: Clinical Science (London, England : 1979)

Article Title: The loss of glycoprotein nonmetastatic melanoma protein B (GPNMB) alters endothelial cell permeability, metabolism, and survival during infectious challenge

doi: 10.1042/CS20256682

Figure Lengend Snippet: (A ) Transendothelial electrical resistance (TEER) of human microvascular endothelial cells (HMVECs) treated with scrambled siRNA (siControl) or GPNMB-targeting siRNA (siGPNMB). ( B ) TEER measurements following lipopolysaccharide (LPS, 100 ng/ml) exposure, normalized to baseline values prior to stimulation. ( C ) TEER measurements after exposure to heat-killed E. coli (HKEC, 3 × 10⁸ cells/ml), normalized to baseline values. ( D-F ) Protein expression levels of ICAM, Integrin-β, and VE-Cadherin, with α-Tubulin used as a loading control. The number of samples used in each experiment ( n ) is expressed in parentheses or dots. Data are presented as mean ± SEM. Statistical analysis: Two-way ANOVA, followed by the Tukey post hoc test.

Article Snippet: Human dermal microvascular endothelial cells (HMVEC) were obtained from the American Type Culture Collection (ATCC CRL-4060; Manassas, VA, U.S.A.).

Techniques: Expressing, Control

Journal: Clinical Science (London, England : 1979)

Article Title: The loss of glycoprotein nonmetastatic melanoma protein B (GPNMB) alters endothelial cell permeability, metabolism, and survival during infectious challenge

doi: 10.1042/CS20256682

Figure Lengend Snippet: Control HMVEC cells (SCR) and treated with siRNA for GPNMB silencing (siGPNMB) were stimulated with vehicle or heat-killed E. coli (HKEC, 3 × 10 8 cells/ml) for 6 h. ( A ) Cell viability (CCK-8 assay) was assessed 1 h after the end of the entire protocol. ( B ) Proliferation (CCK-8 assay) was evaluated at 0-, 12-, and 24 h post-stimulation; the bar graph inserted in the figures represents the area under the curve (AUC) analysis of proliferation at 18 hours. ( C ) Cell migration was assessed using the Scratch-Wounding Assay at 2-, 6-, 12-, and 18 h post-stimulation; the bar graph inserted in the figures represents the quantification of the endpoint of the cell migration curve. ( D ) Representative images of the scratch-wounding assay at 0 and 18 h. ( E-G ) Phosphorylation levels of ERK, JNK, and p38 protein expression were normalized to their respective total protein levels. ( H ) PCNA protein expression normalized to α-Tubulin as a loading control. The number of samples used in each experiment ( n ) is expressed in parentheses or dots. Results are expressed as mean ± SEM. Statistics: Two-way ANOVA, followed by the Tukey post hoc test.

Article Snippet: Human dermal microvascular endothelial cells (HMVEC) were obtained from the American Type Culture Collection (ATCC CRL-4060; Manassas, VA, U.S.A.).

Techniques: Control, CCK-8 Assay, Migration, Phospho-proteomics, Expressing

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