Journal: Regenerative Therapy

Article Title: Airway basal stem cell derived extracellular vesicles promote lung repair in chronic obstructive pulmonary disease

doi: 10.1016/j.reth.2026.101068

Figure Lengend Snippet: Characterization of BC-EVs. (a) Representative transmission electron microscopy image of BC-EVs. Scale bar, 100 nm. (b) Size distribution of BC-EVs determined by Nano-Flow Cytometry. (c) Western blotting analysis showing the expression of EV markers TSG101 and CD63, and the negative marker Calnexin. (d) Representative images of HUVECs tube formation after treatment with DMEM, BC-derived EVs, or MSC-derived EVs. Scale bar, 100 μm. (e) Quantification of the number of tubes formed in each group (n = 5 technical replicates). (f) Cell viability of BCs cultured under serum-depleted conditions and treated with DMEM, or increasing concentrations of BC-EVs or MSC-EVs (n = 3 technical replicates). (e) and (f) All data are presented as mean ± SEM. Statistical analysis was performed using a one-way ANOVA with Tukey's multiple comparisons test. ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001; ns, not significant.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were obtained from the American Type Culture Collection (ATCC) and cultured in DMEM supplemented with 10 % fetal bovine serum.

Techniques: Transmission Assay, Electron Microscopy, Flow Cytometry, Western Blot, Expressing, Marker, Derivative Assay, Cell Culture

Journal: bioRxiv

Article Title: Proteomic analysis of extracellular vesicles released from endothelial cells in vitro reveals increased levels of E-selectin and dual specificity phosphatase 7 as a potential marker of TNFα-mediated apoptosis

doi: 10.64898/2026.01.30.702474

Figure Lengend Snippet: HUVEC were adapted to SFM and treated with TNFα (10 ng/ml) or oxLDL (10 µg/ml) for 20 h. DNA fragmentation was examined by staining cells with propidium iodide staining followed by flow cytometric analysis. (A) Cells were examined by forward scatter vs. side scatter plots and single cells were gated to exclude doublets using peak height against fluorescence on the FL-3 channel. (B) Histogram for untreated cells, (C) TNFα-treated cells and (D) oxLDL-treated cells. Gates were set to define three populations of cells: cells in G1 of the cell cycle, cells in G2, and apoptotic cells with fragmented DNA. (E) The percentage of cells in each gate was calculated (n=3, One Way ANOVA, Tukey’s test ± SD) ** p = 0.0025; *** p = 0.0007; * p = 0.0226. (F) Caspase 3/7 activity was measured following 6 h incubation of HUVEC with different stimuli. Percentage change in RLU compared to the untreated control was calculated for each set (n=6, One Way ANOVA, Tukey’s test ± SD) ** p = 0.0018; *** p = 0.0002. (G) Crystal violet staining was used to determine the number of cells following treatment with stimuli for 20 h (n=8, One Way ANOVA, Tukey’s test ± SD) * p = 0.0342.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) (PromoCell, Heidelberg, Germany) were cultured at 37°C under 5 % (v/v) CO 2 in Endothelial Cell Growth Medium MV (PromoCell) containing 5 % (v/v) foetal calf serum (FCS).

Techniques: Staining, Fluorescence, Activity Assay, Incubation, Control

Journal: bioRxiv

Article Title: Proteomic analysis of extracellular vesicles released from endothelial cells in vitro reveals increased levels of E-selectin and dual specificity phosphatase 7 as a potential marker of TNFα-mediated apoptosis

doi: 10.64898/2026.01.30.702474

Figure Lengend Snippet: HUVEC (2.5 × 10 5 ) in 6-well plates were adapted to SFM and treated with TNFα (10 ng/mL) or oxLDL (10 µg/mL) and incubated at 37°C for 6 h. The conditioned media was then removed and centrifuged at 1500 g for 15 min to remove cell debris. The size distributions and concentration of EVs in the media were examined by NTA. Figures show the size distributions of EVs from (A) untreated HUVEC, (B) HUVEC treated with TNFα, (C) HUVEC treated with oxLDL (n=3 ± SD). (D) The concentrations of EVs released in response to different stimuli were also determined using NTA (n=3 ± SD).

Article Snippet: Human umbilical vein endothelial cells (HUVEC) (PromoCell, Heidelberg, Germany) were cultured at 37°C under 5 % (v/v) CO 2 in Endothelial Cell Growth Medium MV (PromoCell) containing 5 % (v/v) foetal calf serum (FCS).

Techniques: Incubation, Concentration Assay

Journal: bioRxiv

Article Title: Proteomic analysis of extracellular vesicles released from endothelial cells in vitro reveals increased levels of E-selectin and dual specificity phosphatase 7 as a potential marker of TNFα-mediated apoptosis

doi: 10.64898/2026.01.30.702474

Figure Lengend Snippet: A) Principal component analysis showed good discrimination between proteins in HUVEC-derived EVs from untreated and TNFα-treated and oxLDL-treated groups. Each experimental point is an average of the technical replicates. B) Volcano plot of DEPs in EVs from TNFα-treated HUVEC compared to proteins in EVs from untreated cells. C) Volcano plot of DEPs in EVs from oxLDL-treated HUVEC compared to proteins in EVs from untreated cells. D) Volcano plot of DEPs in EVs from TNFα-treated HUVEC compared to proteins in EVs from oxLDL-treated cells. Significantly different proteins were determined as those with an adjusted p -value <0.05 and a fold change >2 (red dots). Data are from three biological replicates each measured in triplicate. E) Heat map of the top 10 significant DEPs in EVs from TNFα-treated cells compared to EVs from untreated cells ranked by log2FC. F) Heatmap for the top 10 significant DEPs in EVs from TNFα-treated cells compared to EVs from oxLDL-treated cells ranked by log2FC.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) (PromoCell, Heidelberg, Germany) were cultured at 37°C under 5 % (v/v) CO 2 in Endothelial Cell Growth Medium MV (PromoCell) containing 5 % (v/v) foetal calf serum (FCS).

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: Proteomic analysis of extracellular vesicles released from endothelial cells in vitro reveals increased levels of E-selectin and dual specificity phosphatase 7 as a potential marker of TNFα-mediated apoptosis

doi: 10.64898/2026.01.30.702474

Figure Lengend Snippet:

Article Snippet: Human umbilical vein endothelial cells (HUVEC) (PromoCell, Heidelberg, Germany) were cultured at 37°C under 5 % (v/v) CO 2 in Endothelial Cell Growth Medium MV (PromoCell) containing 5 % (v/v) foetal calf serum (FCS).

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: Proteomic analysis of extracellular vesicles released from endothelial cells in vitro reveals increased levels of E-selectin and dual specificity phosphatase 7 as a potential marker of TNFα-mediated apoptosis

doi: 10.64898/2026.01.30.702474

Figure Lengend Snippet: Proteins that were significantly and differentially expressed in EVs from untreated and TNFα-treated HUVEC compared to EVs from untreated cells were analysed using A) STRING analysis to examine protein-protein interaction networks showing network edges as confidence (line thickness indicates strength of data to support interactions). PANTHER gene ontology overrepresentation test with an FDR of p<0.05 was used to examine enrichment of B) biological process terms and C) cellular component terms associated with significant DEPs in EVs derived from TNFα-treated cells compared to EVs from untreated cells.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) (PromoCell, Heidelberg, Germany) were cultured at 37°C under 5 % (v/v) CO 2 in Endothelial Cell Growth Medium MV (PromoCell) containing 5 % (v/v) foetal calf serum (FCS).

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: Proteomic analysis of extracellular vesicles released from endothelial cells in vitro reveals increased levels of E-selectin and dual specificity phosphatase 7 as a potential marker of TNFα-mediated apoptosis

doi: 10.64898/2026.01.30.702474

Figure Lengend Snippet: Proteins that were significantly and differentially expressed in EVs from TNFα-treated and oxLDL-treated HUVEC were analysed using A) STRING analysis to examine protein-protein interaction networks showing network edges as confidence (line thickness indicates strength of data to support interactions). PANTHER gene ontology overrepresentation test with an FDR of p<0.05 was used to examine enrichment of B) biological process terms and C) cellular component terms associated with significant DEPs in EVs derived from TNFα-treated compared to EVs from oxLDL-treated cells.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) (PromoCell, Heidelberg, Germany) were cultured at 37°C under 5 % (v/v) CO 2 in Endothelial Cell Growth Medium MV (PromoCell) containing 5 % (v/v) foetal calf serum (FCS).

Techniques: Derivative Assay