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. 2024 Jul 10;20(11):3273–3286. doi: 10.4103/NRR.NRR-D-24-00068

Figure 2.

Figure 2

Validation of the necroptosis model and characterization of NSCs and exosomes.

(A) Characterization of NSC neurospheres. The NSCs displayed a typical spherical neurosphere morphology. Scale bars: 500 μm (upper) and 100 μm (lower). (B) Representative immunofluorescence images of SOX2 (green, Alexa Fluor 488) and Nestin (red, Alexa Fluor 594) expression in NSCs. Both neurospheres and individual NSCs expressed SOX2 and Nestin. Cell nuclei were counterstained with DAPI (blue). Scale bar: 100 μm. (C) NSCs were treated with DMSO or TSZ for 8 hours, after which cell death was analyzed by flow cytometry. (D) Exosome morphology as assessed by transmission electron microscopy. The exosomes in both the NSC-control and NSC-TSZ groups exhibited a typical one-sided semi-concave bilayer membrane structure. Scale bars: 200 nm. (E, F) Nanoflow cytometry detection of particle size distribution, concentration, and exosome surface marker expression. DAPI: 4′,6-Diamidino-2-phenylindole; DMSO: dimethyl sulfoxide; NSC: neural stem cell; SOX2: SRY-box transcription factor 2; TSZ: tumor necrosis factor α, SMAC mimetic, zVAD-fmk.