Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal microvascular endothelial cells (HRMECs) were left untreated or stimulated with tumor necrosis factor-α (TNF-α) (5 ng/mL) for 24 h with or without apigenin (10 µg/mL). Protein expression of intercellular adhesion molecule-1 (ICAM-1) (A) and vascular cell adhesion molecule-1 (VCAM-1) (B) was determined by Western blot analysis. Adhesion of fluorescently labeled THP-1 monocytic cells to HRMECs monolayer was quantified (C) . Results are expressed as mean ± standard deviation or standard error of mean from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three groups and two groups, respectively. *p < 0.05 compared with values obtained from untreated cells. #p < 0.05 compared with values obtained from cells treated with TNF-α (RFU = relative fluorescence unit).

Article Snippet: After reaching a confluent monolayer, overnight starved endothelial cells were 1h pretreated with apigenin (Cat No 1227/10, R&D Systems) followed by 5 ng/ml TNF-α treatment for 16 h. Next, 5 x 10 fluorescent-LeukoTracker labelled monocytic THP-1 cells (American Type Culture Collection, Manassas, VA, USA) were added on the top of treated HRMECs monolayer for 45 min. After washing 3 times, the remaining adherent THP-1 cells were lysed and fluorescence was measured using a spectraMax Gemini-XPS (Molecular Devices, CA, USA) with excitation and emission wavelengths of 485 nm and 538 nm, respectively.

Techniques: Expressing, Western Blot, Labeling, Standard Deviation, Fluorescence