Fig. 2

Cavin2-modified exosomes could be uptaken by aging NPCs via Caveolae-dependent endocytosis. a Schematic graph of the preparation for engineering exosomes. b Representative confocal fluorescence micrographs of Young and Aging NPCs, co-cultured with C-Exo. The exosomes were labeled with PKH26 (red) and the cytoskeleton was labeled with FITC (green). Scale bar, 20 μm. c NPCs were cultured under different inhibition conditions. Representative confocal fluorescence micrographs showed the internalization of PKH26-labeled (red) exosomes in these groups. Chlorpromazine: clathrin-mediated endocytosis inhibitor; Wortmannin: inhibitor of the phosphoinositide 3-kinase; Dynasore: dynamin inhibitor; Filipin: caveolae-dependent endocytosis inhibitor. Scale bar, 20 μm. d Fluorescence intensity of PKH26-labeled C-Exo taken up by NPCs under different inhibition conditions by flow cytometry (n = 3/group). e WB analysis of Cavin2 in Young and Aging NPCs. f WB analysis of CD9, CD63, CD81 and Cavin2 in C-Exo and M-Exo. g Fluorescence intensity of PKH26-labeled C-Exo or M-Exo taken up by Young or Aging NPCs by flow cytometry (n = 3/group). h Representative confocal fluorescence micrographs of PKH26-labeled C-Exo or M-Exo (red) taken up by Young or Aging NPCs. The cytoskeleton was labeled with FITC (green). Scale bar, 20 μm. i Representative confocal fluorescence micrographs of lysosomes (green) and exosomes (red) in NPCs co-cultured with exosomes for 1 h, 2 h and 9 h. Scale bar, 20 μm. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000 1, one-way ANOVA with Tukey’s multiple comparisons. Data are presented as the mean ± SD