Journal: PLoS Pathogens

Article Title: Tumor Cell Marker PVRL4 (Nectin 4) Is an Epithelial Cell Receptor for Measles Virus

doi: 10.1371/journal.ppat.1002240

Figure Lengend Snippet: Wild type IC323 MV infects (A) MCF7 (breast), (B) NCI-H358 (lung) adenocarcinoma and (C) CHO-PVRL4 cell lines via the apical and basolateral surface in Transwell filter assays. Cells were cultivated in Transwell permeable filter supports at a density of 7.0×10 5 cells per Transwell filter (24 mm diameter) for 4 days (MCF7 & NCI-H358) or 2 days (CHO-PVRL4). Cells were then infected from either the apical or basolateral side with IC323-EGFP wtMV. At various times post infection fluorescent images were captured. Scale bar = 500 µm.

Article Snippet: MCF7, NCI-H358, and CHO-PVRL4 cells were seeded onto Transwell permeable filter supports (Corning Inc., 0.4 μm pore size, 24 mm diameter) at a density of 7.0×10 5 cells per well for 4 days (MCF7 & NCI-H358) or 2 days (CHO-PVRL4).

Techniques: Infection

Journal: PLoS Pathogens

Article Title: Tumor Cell Marker PVRL4 (Nectin 4) Is an Epithelial Cell Receptor for Measles Virus

doi: 10.1371/journal.ppat.1002240

Figure Lengend Snippet: (A) Breast (MCF7 and MDA-MB-231) and (B) lung (NCI-H358 and A549) cancer cell lines were grown to confluence on glass coverslips and then fixed with paraformaldehye, permeabilzed, and stained with goat-anti human PVRL4 antibodies (yellow). Nuclei were visualized with TO-PRO-3 nuclear stain (cyan). Images were captured on a Zeiss upright confocal microscope and analyzed using Zen 2008 image capture software (Zeiss). Scale bar = 20 µm. (C) Z-sections of MCF7 and NCI-H358 cells stained with PVRL4 (yellow) and TO-PRO-3 (cyan). PVRL4 is localized to both the apical [A] and basolateral [B] surfaces of these cells. White arrowheads indicate the apical expression of PVRL4. (D) Surface biotinylation of MCF7 cells. MCF7 cells were grown for 96 h on transwell filters (24 mm diameter). The cells were incubated with NHS-biotin from either the apical (lanes A) or basolateral (lanes B) side. After lysis, surface proteins were immunoprecipitated with Neutravidin, and immunocomplexes were subjected to SDS-PAGE and Western blot for PVRL4. Glyceraldehyde 3-phosphate (GAPDH) was used as a loading control.

Article Snippet: MCF7, NCI-H358, and CHO-PVRL4 cells were seeded onto Transwell permeable filter supports (Corning Inc., 0.4 μm pore size, 24 mm diameter) at a density of 7.0×10 5 cells per well for 4 days (MCF7 & NCI-H358) or 2 days (CHO-PVRL4).

Techniques: Staining, Microscopy, Software, Expressing, Incubation, Lysis, Immunoprecipitation, SDS Page, Western Blot, Control

Journal: PLoS Pathogens

Article Title: Tumor Cell Marker PVRL4 (Nectin 4) Is an Epithelial Cell Receptor for Measles Virus

doi: 10.1371/journal.ppat.1002240

Figure Lengend Snippet: MCF7 cells grown on glass coverslips were incubated with 10 µg/ml goat IgG (A,B) or goat anti-PVRL4 (C,D) for 30 min prior to, and during 1 hr adsorption with IC323-EGFP MV via the apical surface. Fluorescence and syncytia formation due to viral infection at 48 hrs was inhibited by the PVRL4 antibody treatment. To determine whether PVRL4 antibodies would also inhibit MV infections via the basolateral route, MCF7 cells were grown on Transwell permeable filter supports as described in . Cells were incubated with 25 µg/ml goat IgG on the apical (E,F,G,H) or basal (I,J,K,L) surface with antibodies specific for human PVRL4 or non-immune antibodies (IgG) for 30 min. Cells were subsequently inoculated with IC323-EGFP MV (MOI = 10) for 4 hrs, also in the presence of antibody. Infections were allowed to proceed for 72 hrs and cells were viewed by fluorescence and bright field microscopy. The interaction of goat polyclonal antibodies with PVRL4 blocked MV infection of MCF7 cells via either the apical or basal routes. Scale bar = 100 µm.

Article Snippet: MCF7, NCI-H358, and CHO-PVRL4 cells were seeded onto Transwell permeable filter supports (Corning Inc., 0.4 μm pore size, 24 mm diameter) at a density of 7.0×10 5 cells per well for 4 days (MCF7 & NCI-H358) or 2 days (CHO-PVRL4).

Techniques: Incubation, Adsorption, Fluorescence, Infection, Microscopy

Journal: Scientific Reports

Article Title: Role of endothelial colony forming cells (ECFCs) Tetrahydrobiopterin (BH4) in determining ECFCs functionality in coronary artery disease (CAD) patients

doi: 10.1038/s41598-022-06758-8

Figure Lengend Snippet: Effect of SIN-1 on ECFC’s migration . (A , B ) represents transwell migrated cells with DAPI nuclear staining; ( A ) Untreated; ( B ) SIN-1 treated. Dot plot ( C ) represents number of migrated cells in lower chamber in untreated and SIN-1 treated groups with individual data points and error bars (n = 3 for each group). Significance was assessed by student ttest and data is represented as mean ± SE. Error bar represent standard error of mean; dots represent average values of the parameter obtained from experimental duplicates p < 0.05 was considered statistically significant.

Article Snippet: Briefly, EGM-2 medium supplemented with 10% FBS was added into the lower chamber, while 5 × 10 4 cells in endothelial basal media (EBM-2) were seeded onto upper chamber of transwell permeable supports (8 μm pore size, polycarbonate membrane, SPL Life Sciences).

Techniques: Migration, Staining