Fig. 5

Knockdown of Notch1 signaling impeded the EVs-mediated effects on NP cells. A NP cells were transfected with siRNA targeted for Notch1 (si-Notch1) or a scrambled siRNA (si-scr), and then treated with EVs (50 μg/ml) for 24 h. The expression levels of Notch1, Hey1, Hey2, ACAN, COL2A1, MMP3 and MMP13 were measured and quantified. B Representative images of Notch1 fluorescence in NP cells and the fluorescence intensity. C Representative images of Wound healing assay and the percentages of filled wounded area were quantified. D The cell viability in different group was evaluated by CCK-8 assay. E Representative immunofluorescent images of Vasorin (green) and Notch1 (red) in NP cells. Nuclei were stained with DAPI (blue). The colocalization analysis of Pearson coefficient was quantified (right panel). F Immunoprecipitation (IP) of Notch1 in NP cells treated with or without EVs and blotted for Notch1 and Vasorin. IgG was used as a negative control. G Mechanism schemes showed the EVs-mediated effects on NP cells. Data were presented as mean ± SD of three independent replicates. P > 0.05 (ns, not significant), *P < 0.05, **P < 0.01, and ***P < 0.001