Journal: British journal of cancer

Article Title: Circulating tumour cells and PD-L1-positive small extracellular vesicles: the liquid biopsy combination for prognostic information in patients with metastatic non-small cell lung cancer.

doi: 10.1038/s41416-023-02491-9

Figure Lengend Snippet: Fig. 3 Characterization of sEVs isolated from plasma samples of 53 patients with NSCLC. a Mode and mean size of sEVs. b Mean concentration of sEVs in the 53 samples. c Representative western blots showing the expression of sEV markers (ALIX, TSG101, CD63, CD81, CD9), PD-L1 and HSP70 in lysates of sEVs isolated from four different plasma samples (upper panels). GRP94 was used as a negative control for cellular material contamination, and the total protein extract of the human lung cancer cell line A549 as a positive control for this contamination (lower panel). d Representative distribution of the size of particles isolated from plasma samples of patients with NSCLC obtained by nanoparticle tracking analysis. e Representative transmission electron microscopy image of the isolated sEVs. Scale bar, 100 nm; sEVs small Extracellular Vesicles.

Article Snippet: Isolated sEVs were then incubated with antibodies against universal sEV markers: CD9 (sc-13118, Santa Cruz Biotechnology), ALIX (NB100-65678, Novus Bio), TSG101 (sc-7964, Santa Cruz Biotechnology), CD63 (NBP2-4225, BioTechne), CD81 (sc166028, Santa Cruz Biotechnology), HSP70 (ADI-SPA810, Enzo Life science), PD-L1 (sc-50298, Santa Cruz Biotechnology), and GRP94 as negative control (ADI-SPA-850, Enzo Life Sciences).

Techniques: Isolation, Clinical Proteomics, Concentration Assay, Western Blot, Expressing, Negative Control, Positive Control, Transmission Assay, Electron Microscopy

Journal: Journal of neuroscience methods

Article Title: Isolation, culture, and characterization of primary endothelial cells and pericytes from mouse sciatic nerve.

doi: 10.1016/j.jneumeth.2025.110366

Figure Lengend Snippet: Fig. 7. Characterized SNEC-EVs and SNPC-EVs promotes nerve regeneration. (A) Representative western blots for EV positive markers (Alix, CD63 and CD81) or EV negative marker (GM130) in SNEC lysate, SNPC lysate, SNEC-EVs, and SNPC-EVs. (B) Neurofilament (green) immunofluorescence staining in mouse DRGs treated with PBS, SNEC-EVs (1 μg/mL and 10 μg/mL), and SNPC-EVs (1 μg/mL and 10 μg/mL) for four days. Scale bar, 100 μm. (C) Quantitative analysis of sprouted neurite length using an image analyzer. Results are presented as means ± SEM (n = 4; *P < 0.05, **P < 0.01, ***P < 0.001). The relative ratio of the PBS group was defined as 1. SNEC, sciatic nerve endothelial cells; SNPC, sciatic nerve pericytes; EVs, extracellular vesicles; DRG, dorsal root ganglion.

Article Snippet: After blocking with 5 % nonfat dry milk for 1 hour at room temperature, membranes were probed with antibodies against the positive EV markers Alix (1:500; Novus Biologicals, Littleton, CO,USA), CD63 (1:500; Novus Biologicals, Littleton, CO, USA), and CD81 (1:500; Novus Biologicals), and the negative EV marker GM130 (BD Biosciences, San Jose, CA, USA) at 4°C overnight.

Techniques: Western Blot, Marker, Immunofluorescence, Staining

Journal:

Article Title: Spirodiketopiperazine-Based CCR5 Inhibitor Which Preserves CC-Chemokine/CCR5 Interactions and Exerts Potent Activity against R5 Human Immunodeficiency Virus Type 1 In Vitro

doi: 10.1128/JVI.78.16.8654-8662.2004

Figure Lengend Snippet: AK602 allows RANTES-induced chemotaxis and CCR5 internalization. (A) CCR5+ MOLT4 cells were exposed to various concentrations of AK530, AK602, E921/TAK-779, or AK671/SCH-C, thoroughly washed, plated onto the upper chamber of the ChemTx System, exposed to 0.5 nM RANTES contained in the lower chamber, and incubated for 4 h; the number of the cells which migrated to the lower chamber was determined, and chemotaxis was calculated. (B) CCR5+ CHO cells were exposed to 10 nM RANTES in the presence or absence of various concentrations of each CCR5 inhibitor and washed with acidic solution for removal of the cell-bound RANTES (21). The amount of cell surface CCR5 was subsequently determined with monoclonal antibody 3A9 (BD PharMingen), which recognizes the N terminus of CCR5 and competes with none of the CCR5 inhibitors tested. In panel A, the level of chemotaxis suppression by TAK-779 and SCH C was greater than that by AK530 and AK602 at four concentrations examined, although complete suppression was seen only at the highest concentration of the AK compounds, 1 μM. However, in panel B, the level of CCR5 internalization suppression by TAK-779 and SCH-C was greater than that of the AK compounds at all three concentrations examined.

Article Snippet: The amount of cell surface CCR5 was subsequently determined with monoclonal antibody 3A9 (BD PharMingen), which recognizes the N terminus of CCR5 and competes with none of the CCR5 inhibitors tested.

Techniques: Chemotaxis Assay, Incubation, Concentration Assay