Fig. 6

NAMPT derived from ApoVs promote biological function of HUVECs via the NAMPT/SIRT1/FOXO1 axis. (A) Western blotting revealed that NAMPT and SIRT1 increased while AC-FOXO1 and FOXO1 decreased in HUVECs after incubation with ApoVs (0, 5, 10, or 20 µg/mL). (B) NAD⁺/NADH ratio in HUVECs after incubation with ApoVs (0, 5, 10, or 20 µg/mL) (n = 3; **P < 0.01 vs. 0 µg/mL). (C) Representative image of FOXO1 immunofluorescence staining in HUVECs. Scale bars, 100 μm. (D) qRT‒PCR results of NAMPT after treating HUVECs with ApoVs. (E) Western blotting for detecting NAMPT in ApoVs, sh-NC-ApoVs and sh-NAMPT-ApoVs. (F) NAD+/NADH ratios in the Ctrl, ApoVs and sh-NAMPT-ApoVs groups. (G) Representative images of EdU, Transwell, wound healing and tube formation assays in the Ctrl, ApoVs and sh-NAMPT-ApoVs groups. (H) Western blotting of NAMPT, SIRT1, AC-FOXO1 and FOXO1 in the Ctrl, ApoVs and sh-NAMPT-ApoVs groups. (I) Western blot analysis of FOXO1 in the cytoplasm (Cyt.) and nucleus (Nuc.) in Ctrl, ApoVs and sh-NAMPT-ApoVs. (J) Representative image of FOXO1 immunofluorescence staining in the Ctrl, ApoVs and sh-NAMPT-ApoVs groups. Scale bars, 100 μm. (K) Quantitative analysis of EdU, Transwell, wound healing, and tube formation assays in (G). All the data are representative of three independent experiments and are shown as means ± SEMs (n = 3; **P < 0.01; ***P < 0.001; ns P > 0.05)