Journal: Frontiers in Molecular Biosciences

Article Title: Molecular determinants of the interaction between HSV-1 glycoprotein D and heparan sulfate

doi: 10.3389/fmolb.2022.1043713

Figure Lengend Snippet: Competition SPR with chemically de-sulfated heparin and sulfated heparosan oligosaccharides reveals 6- O -sulfation as key to gD-HS interaction. (A) Scheme of competition SPR with de-sulfated (deS) heparin ligands, where binding reflects how important the absent sulfate group is to gD/HS interaction. (B) Normalized, average binding percentage of 0.1 µM gD285 injected onto a heparin-immobilized chip pre-mixed with various de-sulfated (+deNS, +de2S, +de6S) heparin oligosaccharides at 1:1 and 1:2.5 M ratios. No oligosaccharide (gD285) and unmodified heparin were used as negative and positive controls, respectively. (C) Normalized, average binding percentage of 0.1 µM gD285 pre-mixed with various sulfated heparosan oligosaccharides (+NS, +NS2S, +NS6S, +NS6S2S) at 1:1 and 1:2.5 M ratios injected over a heparin immobilized SPR chip. Error bars denote the SD of three replicate flow channels.

Article Snippet: The mean fluorescence intensities of the 50 μM glycan concentration of oligosaccharide were plotted against each oligosaccharide identity using GraphPad v. 9.3.1.

Techniques: Binding Assay, Injection

Journal: Journal of Advanced Research

Article Title: Airway basal stem cell-derived extracellular vesicles drive ECM remodeling and suppress fibroblasts activation via the miR-30a-5p/FAP axis in benign tracheal stenosis

doi: 10.1016/j.jare.2025.08.014

Figure Lengend Snippet: Quantification, characterization, and internalization assays of BSC-EVs. (a) Representative fluorescent immunocytochemistry images showing KRT5 (green), MUC5B (green), AC-TUB (green), P63 (red), and KI67 (red) expression in BSCs, with nuclei counterstained using DAPI (blue). Scale bars, 100 μm. (b) HE staining demonstrates the formation of a pseudostratified epithelium by BSCs, comprising ciliated and mucus-producing cells under air–liquid interface differentiation. Scale bar, 30 μm. (c) Nano-flow cytometry analysis reveals the particle size distribution of BSC-EVs. (d) Representative transmission electron microscopy images of BSC-EVs. Scale bar, 300 μm. (e) Western blot analysis confirms the presence of EV markers, including TSG101, HSP70, CD81, CD9, and CD63. #1, #2, #3, #4, and #5 represent BSC-EVs from three different human samples, respectively. (f) Tracking of intratracheally injected PKH-26-labeled BSC-EVs (red) in the rabbit trachea over 14 days, with representative gross, fluorescent stereomicroscope, and fluorescence microscopy images presented sequentially.

Article Snippet: Mean fluorescence intensity was quantified using Celigo software (top right).

Techniques: Immunocytochemistry, Expressing, Staining, Flow Cytometry, Transmission Assay, Electron Microscopy, Western Blot, Injection, Labeling, Fluorescence, Microscopy

Journal: Journal of Advanced Research

Article Title: Airway basal stem cell-derived extracellular vesicles drive ECM remodeling and suppress fibroblasts activation via the miR-30a-5p/FAP axis in benign tracheal stenosis

doi: 10.1016/j.jare.2025.08.014

Figure Lengend Snippet: Assessment of fibroblasts activation and ECM remodeling of miR-30a-5p-containing BSC-EVs in an indirect co-culture system. (a) Schematic representation of the co-culture model for AFs and BSCs transfected with NC or miR-30a-5p, with either DMSO or GW4869 as treatments. (b) Representative fluorescent immunocytochemistry images showing FAP expression in AFs treated with NC inhibitors + DMSO, NC inhibitors + GW4869, miR-30a-5p inhibitors + DMSO, and miR-30a-5p inhibitors + GW4869 (n = 3 per group). Scale bar = 100 μm (top left). Mean fluorescence intensity was quantified using Celigo software (top right). Collagen contractility assays showing representative collagen gel images post-release (bottom left) were quantified for contraction area using ImageJ software (bottom right). (c) Western blot analysis of protein levels for Collagen I, FAP, α-SMA, MMP2, MMP7, MMP9, TIMP1, TIMP2, and TIMP3 in the four treatment groups. (d) Representative images of collagen gels post-release in AFs treated with NC mimics + DMSO, NC mimics + GW4869, miR-30a-5p mimics + DMSO, and miR-30a-5p mimics + GW4869. (e) Corresponding western blot analysis of Collagen I, FAP, α-SMA, MMP2, MMP7, MMP9, TIMP1, TIMP2, and TIMP3 protein levels in these four groups.

Article Snippet: Mean fluorescence intensity was quantified using Celigo software (top right).

Techniques: Activation Assay, Co-Culture Assay, Transfection, Immunocytochemistry, Expressing, Fluorescence, Software, Western Blot