huvecs  (ATCC)

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: PLOS One

Article Title: Rosuvastatin protects against oxLDL-induced endothelial cell oxidative stress and attenuates atherosclerotic plaque formation in ApoE -/- mice through the NF-κB pathway

doi: 10.1371/journal.pone.0339967

Figure Lengend Snippet: (A) Effects of different concentrations of rosuvastatin (0.1, 1, 5 and 10 µM) on HUVEC viability for 24 h. (B) Effects of treatment with different concentrations of ox-LDL (50, 100 and 200 µg/mL) for 24 h on HUVEC viability. (C) HUVECs were treated with different concentrations of rosuvastatin and ox-LDL (200 µg/ml) for 24 h. * P < 0.05, *** P < 0.001 by one-way ANOVA. (D) Bar chart showing the signaling pathways enriched with DEGs in the RNA-Seq dataset ( GSE206927 ) of ox-LDL-treated HUVECs according to GO analysis. (E) Bubble chart showing the signaling pathways enriched with DEGs according to KEGG analysis. (F-G) HUVECs stimulated with ox-LDL and different concentrations of rosuvastatin were stained with DAPI (blue) and Ki67 (purple). Ki67-positive cells were quantified, bar = 100 μm, * P < 0.05, **** P < 0.0001 by one-way ANOVA.

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs) (Cat#PCS-100–010, ATCC, Maryland, USA) were maintained in vascular cell basal medium (Cat#PCS-100–030, ATCC, Maryland, USA) containing ascorbic acid (Cat#PCS-999–006, ATCC, Maryland, USA), FBS (Cat#PCS-999–010, ATCC, Maryland, USA), rhEGF (Cat#PCS-999–018, ATCC, Maryland, USA), heparin sulfate (Cat#PCS-999–011, ATCC, Maryland, USA), L-glutamine (Cat#PCS-999–017, ATCC, Maryland, USA), rhVEGF (Cat#PCS-999–024, ATCC, Maryland, USA), rhFGF-b (Cat#PCS-999–020, ATCC, Maryland, USA), rhIGF-1 (Cat#PCS-999–021, ATCC, Maryland, USA), and hydrocortisone (Cat#PCS-999–014, ATCC, Maryland, USA) at 37°C in an atmosphere containing 5% CO 2 .

Techniques: Protein-Protein interactions, RNA Sequencing, Staining

Journal: PLOS One

Article Title: Rosuvastatin protects against oxLDL-induced endothelial cell oxidative stress and attenuates atherosclerotic plaque formation in ApoE -/- mice through the NF-κB pathway

doi: 10.1371/journal.pone.0339967

Figure Lengend Snippet: HUVECs were treated with ox-LDL in the presence or absence of different concentrations of rosuvastatin (0.1, 1, 5 and 10 µM) for 24 h. (A) NO production in HUVECs. (B) eNOS mRNA expression in HUVECs. (C) A microplate reader was used to measure the fluorescence intensity of the ROS at an excitation wavelength of 488 nm and an absorption wavelength of 525 nm via a fluorescent probe DCFH-DA kit, and Rosup was used as a positive control. (D) The mean intracellular fluorescence intensity was analyzed via fluorescence microscopy. The data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01, ** P < 0.0001 by one-way ANOVA.

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs) (Cat#PCS-100–010, ATCC, Maryland, USA) were maintained in vascular cell basal medium (Cat#PCS-100–030, ATCC, Maryland, USA) containing ascorbic acid (Cat#PCS-999–006, ATCC, Maryland, USA), FBS (Cat#PCS-999–010, ATCC, Maryland, USA), rhEGF (Cat#PCS-999–018, ATCC, Maryland, USA), heparin sulfate (Cat#PCS-999–011, ATCC, Maryland, USA), L-glutamine (Cat#PCS-999–017, ATCC, Maryland, USA), rhVEGF (Cat#PCS-999–024, ATCC, Maryland, USA), rhFGF-b (Cat#PCS-999–020, ATCC, Maryland, USA), rhIGF-1 (Cat#PCS-999–021, ATCC, Maryland, USA), and hydrocortisone (Cat#PCS-999–014, ATCC, Maryland, USA) at 37°C in an atmosphere containing 5% CO 2 .

Techniques: Expressing, Fluorescence, Positive Control, Microscopy

Journal: PLOS One

Article Title: Rosuvastatin protects against oxLDL-induced endothelial cell oxidative stress and attenuates atherosclerotic plaque formation in ApoE -/- mice through the NF-κB pathway

doi: 10.1371/journal.pone.0339967

Figure Lengend Snippet: (A-B) Bcl2 and Bax mRNA expression in HUVECs treated with different concentrations of rosuvastatin and ox-LDL (200 µg/ml) for 24 h. ** P < 0.01, *** P < 0.001, **** P < 0.0001 by one-way ANOVA. (C-D) BCL-2 and Bax protein expression in HUVECs treated with different concentrations of rosuvastatin and ox-LDL (200 µg/ml) for 24 h. The data are presented as the means ± SEMs. * P < 0.05 by one-way ANOVA. (E) HUVECs stimulated with ox-LDL and different concentrations of rosuvastatin were stained with DAPI (blue), Bax (green) and mitochondria (red); scale bar = 20 μm. (F) Early and late apoptosis of HUVECs treated with 100 µg/mL ox-LDL and 10 μmol/L rosuvastatin for 24 h. The quantification results are shown on the right (n = 5). *** P < 0.001, **** P < 0.0001 by one-way ANOVA.

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs) (Cat#PCS-100–010, ATCC, Maryland, USA) were maintained in vascular cell basal medium (Cat#PCS-100–030, ATCC, Maryland, USA) containing ascorbic acid (Cat#PCS-999–006, ATCC, Maryland, USA), FBS (Cat#PCS-999–010, ATCC, Maryland, USA), rhEGF (Cat#PCS-999–018, ATCC, Maryland, USA), heparin sulfate (Cat#PCS-999–011, ATCC, Maryland, USA), L-glutamine (Cat#PCS-999–017, ATCC, Maryland, USA), rhVEGF (Cat#PCS-999–024, ATCC, Maryland, USA), rhFGF-b (Cat#PCS-999–020, ATCC, Maryland, USA), rhIGF-1 (Cat#PCS-999–021, ATCC, Maryland, USA), and hydrocortisone (Cat#PCS-999–014, ATCC, Maryland, USA) at 37°C in an atmosphere containing 5% CO 2 .

Techniques: Expressing, Staining

Journal: PLOS One

Article Title: Rosuvastatin protects against oxLDL-induced endothelial cell oxidative stress and attenuates atherosclerotic plaque formation in ApoE -/- mice through the NF-κB pathway

doi: 10.1371/journal.pone.0339967

Figure Lengend Snippet: (A-D) Protein levels of IkBα, p-IkBα, P65 and p-P65 in HUVECs treated with or without 10 µM rosuvastatin and treated with 100 µg/mL ox-LDL for 24 h. The data are presented as the means ± SEMs. * P < 0.05, ** P < 0.01 by one-way ANOVA. (E) Schematic diagram illustrating the role of rosuvastatin in ox-LDL-induced endothelial cell dysfunction. Rosuvastatin regulates oxidative stress and apoptosis-related gene transcription in endothelial cells by inhibiting ox-LDL-induced IKBα and P65 activation in endothelial cells.

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs) (Cat#PCS-100–010, ATCC, Maryland, USA) were maintained in vascular cell basal medium (Cat#PCS-100–030, ATCC, Maryland, USA) containing ascorbic acid (Cat#PCS-999–006, ATCC, Maryland, USA), FBS (Cat#PCS-999–010, ATCC, Maryland, USA), rhEGF (Cat#PCS-999–018, ATCC, Maryland, USA), heparin sulfate (Cat#PCS-999–011, ATCC, Maryland, USA), L-glutamine (Cat#PCS-999–017, ATCC, Maryland, USA), rhVEGF (Cat#PCS-999–024, ATCC, Maryland, USA), rhFGF-b (Cat#PCS-999–020, ATCC, Maryland, USA), rhIGF-1 (Cat#PCS-999–021, ATCC, Maryland, USA), and hydrocortisone (Cat#PCS-999–014, ATCC, Maryland, USA) at 37°C in an atmosphere containing 5% CO 2 .

Techniques: Activation Assay

Journal: iScience

Article Title: SHP2 regulates VEGFR2 Y1175/PLCγ signaling to impair tumor endothelial barrier stability

doi: 10.1016/j.isci.2026.114784

Figure Lengend Snippet: SHP2 is identified as a PLCγ-dependent VEGFR2 pY1175 interaction partner in endothelial cells (A) Representative western blots showing VEGFR2 immunoprecipitated with antibodies against RASA1, CSK, VAV2, or SHP2 in HUVECs unstimulated (−) or stimulated (+) with VEGFA (100 ng/mL, 5 min). Corresponding whole-cell lysates analyzed using antibodies against phosphorylated VEGFR2 (pY1175), total VEGFR2, and GAPDH as loading control. (B) Quantification of VEGFR2 interaction with RASA1, CSK, VAV2, and SHP2 from (A); n = 3 independent experiments. (C) Representative images of immunostainings for VEC (magenta), pVEC Y685 (green), and DAPI (blue) of unstimulated or VEGFA-stimulated (100 ng/mL, 5 min) HUVECs, pretreated with siCtr or siPTPN11. Scale bars: 30 μm. (D and E) Quantifications of MFI from (C), displayed as fold change relative to unstimulated control. (D) MFI of the VEC area. (E) MFI of pVEC Y685; n = 3 independent experiments, ≥3 fields of view per experiment. (F) Representative images of immunostainings for VEC (magenta), pVEC Y685 (green), and DAPI (blue) of unstimulated or VEGFA-stimulated (100 ng/mL, 5 min) HUVECs, pretreated with siCSK or siVAV2. Scale bars: 30 μm. (G and H) Quantifications of MFI from (F), shown as fold change over unstimulated control; n = 3 independent experiments, ≥3 fields of view per experiment. (I) Representative western blot showing VEGFR2 immunoprecipitated with antibodies against SHP2 or IgG control from unstimulated or VEGFA-stimulated (100 ng/mL, 5 min) HUVECs, pre-treated with siCtr or siPLCG1 . Corresponding whole-cell lysates analyzed by blotting with antibodies against pVEGFR2 Y1175, VEGFR2, PLCγ, and GAPDH as loading control. (J) Quantification of VEGFR2-SHP2 binding from (I); n = 3 independent experiments. (K) Representative western blot showing downstream VEGFA-induced signaling in unstimulated (−) or 100 ng/mL VEGFA-stimulated HUVECs, for 2, 5, 10, and 20 min pretreated with siCtr or siPTPN11. (L–N) Quantification of western blots from (K), shown as fold change relative to unstimulated control. (L) Quantification of western blots for peNOS S1177. (M) Quantification of western blots for pSFK Y418. (N) Quantification of western blots for pVEGFR2 Y1175; n = 4–5 independent experiments. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. HUVECs, human umbilical vein endothelial cells; VEC, VE-cadherin; MFI, mean fluorescence intensity. See also .

Article Snippet: Human Umbilical Vein Endothelial Cells (HUVEC) , PromoCell , C-12203.

Techniques: Western Blot, Immunoprecipitation, Control, Binding Assay, Fluorescence

Journal: iScience

Article Title: SHP2 regulates VEGFR2 Y1175/PLCγ signaling to impair tumor endothelial barrier stability

doi: 10.1016/j.isci.2026.114784

Figure Lengend Snippet: Endothelial PLCγ/SHP2 signaling mediates activation of Src by regulation of both the activating and inhibitory tyrosine phosphorylation sites (A) PLA using antibodies against Src and pSFK Y418 to detect phosphorylation of Src on Y418 in unstimulated or VEGFA-stimulated HUVECs (100 ng/mL, 5min), pretreated with siCtr or siPTPN11 . Endothelial junctions are stained for VEC (magenta) and nuclei with DAPI (blue). Scale bars: 30 μm. Boxed regions in the upper panels are shown at higher magnification in panels below. Scale bars: 10 μm. (B and C) MFI quantifications from (A), displayed as fold change relative to unstimulated control. (B) MFI of the total PLA signal. (C) MFI of the junctional PLA signals representing Y418 phosphorylation of Src; n = 7 independent experiments, ≥3 fields of view per experiment. (D) Representative western blot showing pSFK Y529 signaling in unstimulated (−) or VEGFA-stimulated HUVECs (100 ng/mL) for 2, 5, 10 and 20 min, pretreated with siCtr or si PTPN11 . (E) Quantification of western blots from (D); n = 5 independent experiments. (F and G) PLA using antibodies against Src and pSFK 529, visualizing phosphorylation of Src at the inhibitory phosphosite in HUVECs stimulated for 2, 5, and 10 min or left unstimulated. (F) Phosphorylation of Src at the inhibitory phosphosite in HUVECs pre-treated with siCtr . (G) Phosphorylation of Src at the inhibitory phosphosite in HUVECs pre-treated with siPTPN11 . Endothelial junctions are stained for VEC (magenta) and nuclei with DAPI (blue). Scale bars: 30 μm. Boxed regions in the upper panels are shown at a higher magnification in panels below. Scale bars: 10 μm. (H) Quantification of PLA experiments from (F) and (G) and F; n = 6 independent experiments, ≥3 fields of view per experiment. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. PLA, proximity ligation assay; HUVECs, human umbilical vein endothelial cells; VEC, VE-cadherin. See also .

Article Snippet: Human Umbilical Vein Endothelial Cells (HUVEC) , PromoCell , C-12203.

Techniques: Activation Assay, Phospho-proteomics, Staining, Control, Western Blot, Proximity Ligation Assay

Journal: iScience

Article Title: SHP2 regulates VEGFR2 Y1175/PLCγ signaling to impair tumor endothelial barrier stability

doi: 10.1016/j.isci.2026.114784

Figure Lengend Snippet: PLCγ/SHP2 interplay leads to eNOS activation followed by Src nitration (A) Representative western blot showing eNOS T495 signaling in unstimulated (−) or 100 ng/mL VEGFA-stimulated HUVECs for 2, 5, 10 and 20 min, pre-treated with siCtr or siPTPN11 . (B) Quantification of western blots from (A); n = 4 independent experiments. (C) PLA for NitroTyr and pSFK Y418 to detect full activation of Src in HUVECs stimulated with VEGFA (100 ng/mL, 5 min) or left unstimulated, and pretreated with siCtr or siPTPN11 . Endothelial junctions are stained for VEC (magenta) and DAPI (blue). Scale bars: 30 μm. Boxed regions in the upper panels are shown at higher magnification in panels below. Scale bars: 10 μm. (D) Quantification of junctional MFI PLA signals representing Y418 phosphorylation and 3-nitration of Src from (C), displayed as fold change to unstimulated control; n = 5 independent experiments, ≥3 fields of view per experiment. (E) Western blot showing eNOS S1177 and SHP2 Y542 signaling in unstimulated (−) or 100 ng/mL VEGFA-stimulated HUVECs for 2, 5, 10, and 20 min, pre-treated with siCtr or siPLCG1 . (F and G) Quantifications of western blots from (E). (F) Quantifications of western blots for peNOS S1177. (G) Quantifications of western blots for pSHP2 Y542; n = 4 independent experiments. (H) Representative immunostaining images with antibodies against VE-cadherin (VEC; magenta) and pSHP2 Y542 (green), in HUVECs unstimulated or stimulated with VEGFA (100 ng/mL, 5min) after downregulation with siCtr or siPLCG1 . Scale bars: 30 μm. (I) Quantification of MFI from (H), shown as fold change relative to unstimulated control; n = 4 independent experiments, ≥3 fields of view/experiment. One-way ANOVA. Data represent the mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001. VEC, VE-cadherin. See also .

Article Snippet: Human Umbilical Vein Endothelial Cells (HUVEC) , PromoCell , C-12203.

Techniques: Activation Assay, Nitration, Western Blot, Staining, Phospho-proteomics, Control, Immunostaining