Journal: Materials Today Bio

Article Title: Modulation of endothelial-to-mesenchymal transition via NRP-1 targeting with melittin attenuates pulmonary fibrosis

doi: 10.1016/j.mtbio.2025.102659

Figure Lengend Snippet: MLT attenuates TGF-β1-induced EndMT by inhibiting the TGF-β/Smad and MAPK pathways in HUVECs. (A–D) The mRNA levels of endothelial (CD31, VE-cadherin) and mesenchymal (α-SMA, vimentin) markers were determined by RT-qPCR. (E) EndMT-related protein expression of CD31, VE-cadherin, α-SMA, vimentin, Twist, and Snail was examined by western blot. The original western blot images are provided in Quantitative analysis of CD31, VE-cadherin, α-SMA, vimentin, Twist, and Snail protein levels (relative to β-actin). (L) Immunofluorescence staining of EndMT markers in HUVECs. Scale bar: 50 μm; Blue: DAPI; Green: CD31; Magenta: VE-cadherin; Red: α-SMA; Orange: vimentin. (M) Nuclear transcription factor immunofluorescence during EndMT in HUVECs. Scale bar: 50 μm; Blue: DAPI; Green: Twist; Red: Snail. (N) Protein levels of key components in the TGF-β/Smad and MAPK pathways were assessed by western blot. The original western blot images are provided in Quantitative analysis of Smad4/β-actin, p-Smad2/3/Smad2/3, p-ERK/ERK, p-JNK/JNK and p-P38/P38 protein levels. Data are presented as the mean ± SEM (n = 3); ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Antibodies: VE-cadherin, β-actin and HRP-labeled antibodies were purchased from Boster Company.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Staining

Journal: Materials Today Bio

Article Title: Modulation of endothelial-to-mesenchymal transition via NRP-1 targeting with melittin attenuates pulmonary fibrosis

doi: 10.1016/j.mtbio.2025.102659

Figure Lengend Snippet: Nanoparticle delivery of MLT potentiates anti-PF efficacy. (A) Experimental timeline of BLM-induced PF and therapeutic intervention. Mice received intratracheal BLM on day 0, followed by intraperitoneal administration of pLNPs, MLT-L (0.5 mg/kg), MLT-H (2 mg/kg), M-pLNPs (0.5 mg/kg as MLT) every other day from day 7 to day 21. (B) Representative macroscopic morphological images of lung tissues from each treatment group. (C) Histopathological analyses of lung tissues. Scale bar: 20 μm. (D) Quantitative analysis of fibrosis severity by Ashcroft scoring. (E) Quantitative assessment of Masson staining. (F) Representative Micro-CT images of lung tissue at three anatomical planes: sagittal, coronal, and transverse, demonstrating the spatial distribution of fibrotic lesions. Scale bar: 15 mm. (G) Immunofluorescence staining of key therapeutic targets in lung tissues. Scale bar: 50 μm. Blue: DAPI; Red: CD31; Yellow: VE-cadherin; Magenta: α-SMA; Green: COL-I; Cyan: Vimentin. (H) Immunofluorescence staining images showing the expression and localization of nuclear transcription factors Twist and Snail in lung tissues. Scale bar: 50 μm. Blue: DAPI; Red: Twist; Magenta: Snail. (I–N) Quantitative analysis of pulmonary function parameters including respiratory system resistance (Rrs) (I), elastance (Ers) (J), compliance (Crs) (K), forced vital capacity (FVC) (L) and forced expiratory volume at 0.2 s (FEV0.2) (M), and static compliance (Cst) (N). Data are presented as the mean ± SEM (n = 5); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, ns: not significant.

Article Snippet: Antibodies: VE-cadherin, β-actin and HRP-labeled antibodies were purchased from Boster Company.

Techniques: Staining, Micro-CT, Immunofluorescence, Biomarker Discovery, Expressing

Journal: Frontiers in Pharmacology

Article Title: Positive Effect of Andrographolide Induced Autophagy on Random-Pattern Skin Flaps Survival

doi: 10.3389/fphar.2021.653035

Figure Lengend Snippet: The upregulation of angiogenesis in the skin flaps via Andro (A,C) IHC of Cadherin5 and VEGF in both groups of ischemic skin flaps (original magnification, ×200; scan bar, 50 μm). (B,D) The total absorbance of Cadherin5 and VEGF in IHC. (E) The results of immunoblotting i.e., the expressions of cadherin 5, MMP9, VEGF in the control as well as Andro groups. (F–H) The quantification of Cadherin5, MMP9, and VEGF expressions in the flaps by measuring their optical densities. The obtained data were presented as means ± SEM. Significance: * p -value < 0.05 and ** p -value < 0.01, vs. control group (n = 6 per group).

Article Snippet: Diaminobenzidine (DAB) developer, pentobarbital sodium, and the H&E Kit were procured from Solarbio Science and Technology (Beijing, China), while Cdh5-Rabbit Monoclonal Antibodies were provided via Boster Biological Technology, China.

Techniques: Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Positive Effect of Andrographolide Induced Autophagy on Random-Pattern Skin Flaps Survival

doi: 10.3389/fphar.2021.653035

Figure Lengend Snippet: 3MA reversed the effects of Andro on random-pattern skin flap viability (A) The immunofluorescence data of LC3II in the ischemic skin flap (scan bar, 50 μm). (B) T he histogram representing the percentage of the LC3II-positive cells. (C,E) The immunoblotting of the autophagy associated proteins VPS34, p62, LC3II, Beclin 1, and CTSD; angiogenesis associated proteins VEGF, cadherin5, and MMP9; the oxidative stress-related proteins i.e., SOD1, HO1, and eNOS and the apoptosis-associated proteins i.e., Bax, CYC, and CASP3. (D,F) The quantification of autophagy-, angiogenesis, and oxidative stress-related proteins by measuring their optical densities, as previously mentioned. The obtained data were presented as means ± SEM. Significance: * p -value < 0.05 and ** p -value < 0.01, vs. control group (n = 6 per group).

Article Snippet: Diaminobenzidine (DAB) developer, pentobarbital sodium, and the H&E Kit were procured from Solarbio Science and Technology (Beijing, China), while Cdh5-Rabbit Monoclonal Antibodies were provided via Boster Biological Technology, China.

Techniques: Immunofluorescence, Western Blot, Control

Journal:

Article Title: Transference of recombinant VE-cadherin cytoplasmic domain alters endothelial junctional integrity and porcine microvascular permeability

doi: 10.1113/jphysiol.2003.051086

Figure Lengend Snippet: A, molecular structure of VE-cadherin. EC1–EC5, extracellular domain; TM, transmembrane region; CPD, cytoplasmic domain. Black region represents the signal peptide. Grey boxes indicate the binding regions of p120 and β/γ catenins, respectively. B, schematic composition of the expression vector construct pQE30/VE-cad-CPD. PT5, phage promoter T5; lacO, lac operator; RBS, ribosome binding site; ATG, start codon; 6xHis, histidine tag. C, cDNA product of VE-cadherin cytoplasmic domain on agarose gel stained with ethidium bromide. D, Western blot detection of the rVE-cad CPD protein. Lane 1, purified protein; lane 2, mock extract from E. coli without IPTG induction; lanes 3–5, cell lysate from different bacterial clones.

Article Snippet: For the determination of the endogenously expressed VE-cadherin distribution, cells were incubated with a monoclonal antibody to the VE-cadherin extracellular domain (BD Bioscience) and followed by a FITC-labelled antimouse conjugate.

Techniques: Binding Assay, Expressing, Plasmid Preparation, Construct, Agarose Gel Electrophoresis, Staining, Western Blot, Purification, Clone Assay

Journal:

Article Title: Transference of recombinant VE-cadherin cytoplasmic domain alters endothelial junctional integrity and porcine microvascular permeability

doi: 10.1113/jphysiol.2003.051086

Figure Lengend Snippet: Cells were transfected with mock extract (A and C) or rVE-cad CPD (B and D) and then subjected to immunolabelling with a monoclonal antibody directed to either the extracellular domain of VE-cadherin or β-catenin, followed by a FITC-conjugated secondary antibody. VE-cadherin staining changed from a uniform distribution along the cell membrane under control conditions (A) to a diffuse pattern after rVE-cad CPD treatment. β-Catenin staining was mainly localized at cell–cell contact areas, with control cells showing a continuous distribution in contrast to the discontinuous, stitch-like pattern in transfected cells. Note the stitch-like structures and adjacent intercellular holes (arrows).

Article Snippet: For the determination of the endogenously expressed VE-cadherin distribution, cells were incubated with a monoclonal antibody to the VE-cadherin extracellular domain (BD Bioscience) and followed by a FITC-labelled antimouse conjugate.

Techniques: Transfection, Staining

Journal:

Article Title: Transference of recombinant VE-cadherin cytoplasmic domain alters endothelial junctional integrity and porcine microvascular permeability

doi: 10.1113/jphysiol.2003.051086

Figure Lengend Snippet: Endothelial cells were lysed and immunoprecipitated (IP) with protein G PLUS-agarose preabsorbed with a goat polyclonal antibody to β-catenin. The immunoprecipitates were separated by SDS-PAGE on 4–12% Tris-glycine gels, transferred to nitrocellulose membranes, and blotted with a monoclonal antibody to either β-catenin (A), VE-cadherin (B) or rVE-cad CPD (C). For the detection of rVE-cad CPD, the immunoprecipitates were fractionated by SDS-PAGE on 4–20% gel, followed by protein transfer and immunoblotting with an antihistidine antibody. After incubation with a secondary antibody conjugated to horseradish peroxidase, immunoreactive bands were detected using LumiGLO chemiluminescent substrate, scanned by reflectance scanning densitometry, and quantified using NIH Image software. Lane 1, non-treated cells; lane 2, TransIT-LT1 only; lane 3, mock-transfected cells; lane 4, cells transfected with rVE-cad CPD (1 μg ml−1); and lane 5, cells transfected with rVE-cad CPD (5 μg ml−1). Each blot represents four separate experiments. The bar graph under each representative image shows the optical density of the bands. Results are expressed as percentage of control (*P < 0.05versus control).

Article Snippet: For the determination of the endogenously expressed VE-cadherin distribution, cells were incubated with a monoclonal antibody to the VE-cadherin extracellular domain (BD Bioscience) and followed by a FITC-labelled antimouse conjugate.

Techniques: Immunoprecipitation, SDS Page, Western Blot, Incubation, Software, Transfection