Journal: Scientific reports

Article Title: Small extracellular vesicles convey the stress-induced adaptive responses of melanoma cells.

doi: 10.1038/s41598-019-51778-6

Figure Lengend Snippet: Figure 1. Schematic illustration of the experimental workflow in six steps. B16F1 mouse melanoma cell- derived sEVs were isolated and characterised by Western Blot (WB), atomic force microscopy (AFM) and dynamic light scattering (DLS). Then B16F1 cultures were treated in five different ways, and 72 h supernatants were harvested for sEV isolation. Vesicle samples were then analysed by nanoparticle tracking analysis (NTA) to determine the number of released sEVs, sequencing and LC-MS/MS to describe their miRNome and proteome. Ingenuity Pathway Analysis (IPA) was used to analyse data and predict the functional differences between sEV groups. This in silico predictions were tested in vitro on mesenchymal stem cell (MSC) and melanoma cell cultures and MSC-B16F1 3D co-cultures as well using Ki-67-specific immunocytochemistry, Cell-Clock cell cycle assay, wound healing assay, and 3D hanging drop technology. Abbreviation: n.ctrl-negative control. Figure was created with BioRender.com.

Article Snippet: Purity of MSC cultures was checked by flow cytometry using the Mouse Multipotent Mesenchymal Stromal Cell Marker Antibody Panel (R&D Systems) according to the manufacturer’s instructions and a FACSCalibur instrument coupled with CellQuest Pro 6.0 (BD Biosciences).

Techniques: Derivative Assay, Isolation, Western Blot, Microscopy, Sequencing, Liquid Chromatography with Mass Spectroscopy, Functional Assay, In Silico, In Vitro, Immunocytochemistry, Cell Cycle Assay, Wound Healing Assay, Negative Control