Journal: Frontiers in Immunology

Article Title: CMTM3 regulates vascular endothelial cell dysfunction by influencing pulmonary vascular endothelial permeability and inflammation in ARDS

doi: 10.3389/fimmu.2025.1544610

Figure Lengend Snippet: Mice, cell lines, and reagents.

Article Snippet: Lipopolysaccharide (HUVECs) , Solarbio , IL2020.

Techniques: Over Expression, Knockdown, Cell Culture, Staining, cDNA Synthesis, SYBR Green Assay, Real-time Polymerase Chain Reaction, Membrane

Journal: Frontiers in Immunology

Article Title: CMTM3 regulates vascular endothelial cell dysfunction by influencing pulmonary vascular endothelial permeability and inflammation in ARDS

doi: 10.3389/fimmu.2025.1544610

Figure Lengend Snippet: (A) The expression levels of CMTM3 transcripts in human umbilical vein endothelial cells (HUVECs) treated with 100 ng/ml LPS for 2h, 6h, and 24h ( n ≧ 3). (B) The expression levels of CMTM3 transcripts in HUVECs subjected to stimulation with 4h-hypoxia/2h-reoxygenation and 5h-hypoxia/2h-reoxygenation. Unstimulated HUVECs were used as the control ( n ≧ 3).

Article Snippet: Lipopolysaccharide (HUVECs) , Solarbio , IL2020.

Techniques: Expressing, Control

Journal: Frontiers in Immunology

Article Title: CMTM3 regulates vascular endothelial cell dysfunction by influencing pulmonary vascular endothelial permeability and inflammation in ARDS

doi: 10.3389/fimmu.2025.1544610

Figure Lengend Snippet: CMTM3 overexpression enhances permeability of the human umbilical vein endothelial cells (HUVECs) under in-vitro ADRS inflammatory conditions ( n ≧ 3). (A) CMTM3 expression levels in control HUVECs as well as adsham and adCMTM3-transfected HUVECs. (B) FITC-dextran assay results show the cellular permeability based on relative fluorescence leakage (RFI) in the adsham- and adCMTM3-transfected HUVECs that were untreated or treated with 100 ng/ml LPS for 6h. (C) FITC-Dextran assay results show the cellular permeability based on relative fluorescence leakage (RFI) in the adsham- and adCMTM3-transfected HUVECs that were untreated or treated with 4h-hypoxia/2h-reoxygenation.

Article Snippet: Lipopolysaccharide (HUVECs) , Solarbio , IL2020.

Techniques: Over Expression, Permeability, In Vitro, Expressing, Control, Transfection, Fluorescence

Journal: Frontiers in Immunology

Article Title: CMTM3 regulates vascular endothelial cell dysfunction by influencing pulmonary vascular endothelial permeability and inflammation in ARDS

doi: 10.3389/fimmu.2025.1544610

Figure Lengend Snippet: CMTM3 knockdown reduces human umbilical vein endothelial cell (HUVEC) permeability under in-vitro inflammatory conditions. (A) CMTM3 expression levels in the control, shsham, and shCMTM3 HUVECs. (B) FITC-dextran assay results show the cellular permeability based on relative fluorescence leakage (RFI) in the shsham and shCMTM3-transfected-HUVECs that were untreated or treated with 100 ng/ml LPS for 6h. (C) FITC-dextran assay results show the cellular permeability based on relative fluorescence leakage (RFI) of shsham and shCMTM3-transfected HUVECs that were untreated or treated with 4h-hypoxia/2h-reoxygenation.

Article Snippet: Lipopolysaccharide (HUVECs) , Solarbio , IL2020.

Techniques: Knockdown, Permeability, In Vitro, Expressing, Control, Fluorescence, Transfection

Journal: Frontiers in Immunology

Article Title: CMTM3 regulates vascular endothelial cell dysfunction by influencing pulmonary vascular endothelial permeability and inflammation in ARDS

doi: 10.3389/fimmu.2025.1544610

Figure Lengend Snippet: The expression levels of IL-6 and TNF-α in the adCMTM3- and shCMTM3-transfected HUVECs after (A, C) LPS stimulation for 6 h and (B, D) 4h/2h hypoxia/reoxygenation treatment.

Article Snippet: Lipopolysaccharide (HUVECs) , Solarbio , IL2020.

Techniques: Expressing, Transfection

Journal: Frontiers in Immunology

Article Title: CMTM3 regulates vascular endothelial cell dysfunction by influencing pulmonary vascular endothelial permeability and inflammation in ARDS

doi: 10.3389/fimmu.2025.1544610

Figure Lengend Snippet: RNA sequencing analysis of lipopolysaccharide (LPS)–treated shCMTM3-HUVECs. (A) Top 20 KEGG pathways representing DEGs between shCMTM3-HUVECs and shsham- human umbilical vein endothelial cells (HUVECs). (B) Top 20 KEGG pathways between LPS-treated shCMTM3-HUVECs and LPS-treated shsham-HUVECs. (C) PPI network analysis of DEGs between shCMTM3-HUVECs and shsham-HUVECs. (D) PPI network analysis of DEGs between LPS-treated shCMTM3-HUVECs and LPS-treated shsham -HUVECs.

Article Snippet: Lipopolysaccharide (HUVECs) , Solarbio , IL2020.

Techniques: RNA Sequencing

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: Nogo-B affected HUVECs proliferation and tube formation by transfected with GV248-Nogo-B RNAi-GFP. (A) The cells grew well and shaped like spindles with no vacuole seen in the nucleus and were transfected under a fluorescence microscope and the whole cell emits fluorescence (200X). (B) Detection of the Nogo-B expression by Western Blot. Nogo-B was completely knocked down. (C) The effect of Nogo-B on proliferation and (D) tube-formation of HUVECs were measured by CCk-8 assay and tube formation assay. Data was shown as means ± SD, (C) n=3; (D) n=5; *P<0.05, **P<0.01, ***P<0.001. Scar bars: 100 μm.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Transfection, Fluorescence, Microscopy, Expressing, Western Blot, CCK-8 Assay, Tube Formation Assay

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: Cytokines detected in HUVECs supernatant after knocked down Nogo-B. A. Cytokines in supernatant of HUVEC-NC and HUVEC-siNogo-B detected by antibody microarray. They were labeled by Cy3 and Cy5, which were two different fluorescent molecular. B. It showed the ratio of siNogo-B/NC from antibody microarray. EGF, VEGF-D, FGF-4 and Progranulin were increased obviously in the HUVEC-siNogo-B supernatant, whereas anti-angiogenesis cytokines MMP-19 and VEGI declined.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Microarray, Labeling

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: HUVEC-siNogo-B-CM exhibited chemoattractive, mitogenic effects on fibroblasts. (A, B) The pictures showed the scratch test of fibroblasts in 24 h. The number of migrating fibroblasts in 50% HUVEC-siNogo-B group was significantly greater than that in 50% HUVEC-NC group in 24 h the migration (C) and proliferation (D) of fibroblasts were measured by Transwell Assay and CCk-8 Assay. The proliferative activity of fibroblasts increased markedly in 50% HUVEC-siNogo-B group as compared with 50% HUVEC-NC group after 4 days of co-culturing. Data was shown as means ± SD, n=3, *P<0.05, ***P<0.001. The scale bar is 500 µm.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Migration, Transwell Assay, CCK-8 Assay, Activity Assay

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: The HUVECs-siNogo-B-CM promoted migration of epidermal cells. The migration of epidermal cells was measured by scratch test (A, B) and CCk-8 Assay (C). The epidermal cells in the 50% HUVEC-siNogo-B group was significantly greater than that in the 50% HUVEC-NC group in 12 h and 36 h. The scale bar is 500 µm. The epidermal cells in 50% HUVEC-siNogo-B group increased significantly faster after 4 days co-culturing. 5% FBS was treated as positive control and HUVEC-NC, DMEM as negtive control. Data was shown as means ± SD, n=3; *P<0.05, ***P<0.001. The scale bar is 500 µm.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Migration, CCK-8 Assay, Positive Control, Control

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: HUVEC-siNogo-B could survive in 48 h after injected intradermally around the excisional wound on mice. The flow cytometry and the Immunofluorescence were performed to detected the survival of HUVEC-siNogo-B (GFP+) cells. The blue spots in the top left corner represents alive cells (A). The quantity of blue spots in two injection groups was much higher compared with vehicle control group. (B) Representative immunofluorescent staining of The GFP-labeled HUVECs. GFP+DAPI+ were survival cells by initially injected.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Injection, Flow Cytometry, Immunofluorescence, Control, Staining, Labeling

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: Topical administration of HUVEC-siNogo-B accelerated wound healing on mice. A. The wound was treated with saline, HUVEC-NC and HUVEC-siNogo-B, examined at day 1, day 3, day 5, day 7, day 14 after wounding and digitally photographed. B. Quantitative analysis of wound closure demonstrating higher wound healing rate in the HUVEC-siNogo-B group. Data was shown as means ± SD, n=6; ***P<0.001.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Saline

Journal: American Journal of Translational Research

Article Title: Transplantation of HUVECs with genetically modified Nogo-B accelerates wound-healing in nude mice

doi:

Figure Lengend Snippet: The HUVEC-siNogo-B increased callipary density in wound of nude mice. (A) Representative HE staining at day 7 after wounding. (B) Representative Immunohistochemistry staining of CD31 at day 7. Statistical analysis of CD31+ newly formed vessels showed that neovascularization was more obvious in HUVEC-siNogo-B treated wounds (C). Data was shown as means ± SD, n=5; ***P<0.001. The scale bar is 100 µm.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) were purchased from ScienCell (San Diego, USA) and cultured in endothelial cell medium (ECM, Gibco, Life Technologies) supplemented with 10% FBS.

Techniques: Staining, Immunohistochemistry