Journal: bioRxiv

Article Title: A Biphasic Effect of Alcohol on Endothelial Plasticity Through Regulation of Endothelial-to-Mesenchymal Transition

doi: 10.64898/2026.04.14.718463

Figure Lengend Snippet: (a-c) SM22α, eNOS, αSMA, and Fibronectin (FN) mRNA levels determined by qRT-PCR in human coronary artery endothelial cells (HCAEC) following treatment with TGFβ1 (10 ng/ml, 7d), hypoxia (5% O 2 , 4d) or TGFβ1 plus hypoxia (4d), in the absence or presence of ethanol (EtOH) at concentrations indicated (0-100 mM range). (d) SM22α, eNOS, and Cdh5 mRNA levels in HCAEC treated with TGFβ1 (10 mg/ml, 2d) +/- EtOH 25 mM or 100 mM. (e) Representative western blots showing CD31, Cdh5, SM22α, and SNAIL protein expression in HCAEC treated with TGFβ1, IL-1β, or Hypoxia +/- EtOH (25 mM or 100 mM). β-actin or GAPDH were used as loading controls. Data are mean±SEM, n=3. *p<0.05 vs control (no treatment), #p<0.05 vs TGFβ1 or hypoxia.

Article Snippet: Human coronary artery endothelial cells (HCAEC, C-12221) and Human umbilical vein endothelial cells (HUVEC-pooled, C-12203) were obtained from PromoCell.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: A Biphasic Effect of Alcohol on Endothelial Plasticity Through Regulation of Endothelial-to-Mesenchymal Transition

doi: 10.64898/2026.04.14.718463

Figure Lengend Snippet: (a) (b) Representative images of immunofluorescently stained carotid cross sections from sham-operated and ligated controls, moderate EtOH, and Binge EtOH experimental groups (males). Blue = Dapi nuclear stain, red = Tm-Cdh5+, and white=αSMA+. (c) Cells co-expressing Cdh5 and αSMA (i.e., myo-endothelial cells indicative of EndMT) were quantified using QuPath bioimage analysis software in carotid cross sections post-ligation from controls (grey bars), moderate EtOH (green bars), and binge EtOH (red bars) experimental groups. Bar graphs show cumulative data expressed as % Myoendothelial cells /full cross section, % Myoendothelial cells /neointima, or Number of myoendothelial cells/full cross section. Data are mean±SEM, n=25-30 sections from 5-6 mice). *p<0.05, ** P<0.001, **** p<0.0001.

Article Snippet: Human coronary artery endothelial cells (HCAEC, C-12221) and Human umbilical vein endothelial cells (HUVEC-pooled, C-12203) were obtained from PromoCell.

Techniques: Staining, Expressing, Software, Ligation

Journal: bioRxiv

Article Title: Molecular mechanisms of 10-Butyl Ether Minocycline (BEM), a novel non-antibiotic tetracycline, as a potential treatment for inflammatory and neuroimmune-related disorders

doi: 10.1101/2025.06.23.661183

Figure Lengend Snippet: Human umbilical vein endothelial cells (HUVECs) were seeded at 3 × 10⁴ cells/well in endothelial cell medium into a two-well silicone insert to create a defined cell-free gap. After adherence, cells were treated with 25 ng/mL VEGF (positive control) or no VEGF (negative control), followed by treatment with either minocycline (MINO) or butyl ether minocycline (BEM) at 100 µM. (a) Representative phase-contrast images show VEGF-stimulated migration and its inhibition by BEM and MINO. (b) Quantification of the remaining gap area revealed significant inhibition of VEGF-induced migration by both BEM and MINO. Data are shown as mean ± SEM (n = 4 biological replicates per group). Statistical analysis was performed by two-way ANOVA with Šídák’s post hoc multiple comparisons test. Significance: ***p = 0.0003 (VEGF vs. Control), ***p = 0.0009, (VEGF vs. VEGF + MINO), **p = 0.0022 (VEGF vs. VEGF + BEM).

Article Snippet: Primary HUVEC (Human Umbilical Vein Endothelial Cells) cells (PromoCell, Heidelberg, Germany) were seeded at 3×10 4 cells/well in 70 µL endothelial cell media (PromoCell, Heidelberg, Germany) into a 2-well silicone insert (Ibidi, Munich, Germany) and incubated overnight at 37 °C with 5% CO 2 .

Techniques: Positive Control, Negative Control, Migration, Inhibition, Control

Journal: Clinical Cancer Research

Article Title: Targeting Neuropilin 1 as an Antitumor Strategy in Lung Cancer

doi: 10.1158/1078-0432.ccr-07-0001

Figure Lengend Snippet: Fig. 5. Cyclic 7-mer peptides bind NRP1and inhibit CL1-5 invasion and angiogenesis in vivo. A, the selected peptides reduce phosphorylation ofVEGFR2. HUVECs were pretreated with peptides for10 min followed by treatment withVEGF for 5 min. Phosphorylation ofVEGFR2 was determined byWestern blotting with an anti ^ phospho-VEGFR2 antibody.TotalVEGFR2 was determined byWestern blotting with an anti-VEGFR2 antibody. B, the effect of peptides on CL1-5 cells invasive activity. The invasive activity of cells was detected by invasion assay. CL1-5 cells (2.5 104) were seeded onTranswells coated with 30 Ag matrigel and incubated with peptides DG1or DG2 for 48 h.Then, the cells that had invaded the membrane were counted.Values were normalized to the relative invasion activity of the nontreated control cells. Experiments were done in triplicate, three independent times. In DG1- and DG2-treated cells, the invasion ability was statistically significantly different across the various concentrations of DG1or DG2 byANOVA (DG1, P < 0.001; DG2, P = 0.011). C, the effect of peptides on tumor angiogenesis in vivo. Immunohistochemical staining of the Matrigel plug sections with an anti-CD31antibody showed a significant decrease in CD31-positive vessels in plugs containing DG1peptide compared with mock-treated plugs. Original magnification, 200.The counts of microvessels surrounding the tumor nests were calculated. D, effect of peptides on tumorigenesis in vivo.Volumes of tumors from control CL1-5 cells (E) and DG1-treated cells (n) were measured at the indicated times as described in Materials and Methods. Means and 95% CI are shown (n = 5 mice per group). E, summary diagram showing thatVEGF165 can bind to NRP1and trigger the NRP1/VEGFR2/PI3K/Akt signaling pathways and result in tumor angiogenesis, cancer cell invasion, and tumorigenesis.The synthetic peptides DG1/DG2 can specifically block this signaling pathway and may have therapeutic potential.

Article Snippet: Human umbilical vascular endothelial cells (HUVEC) and culture media were purchased from Cell Applications, Inc.

Techniques: In Vivo, Phospho-proteomics, Activity Assay, Invasion Assay, Incubation, Membrane, Control, Immunohistochemical staining, Staining, Protein-Protein interactions, Blocking Assay