Extended Data Fig. 1: Characterization of SF1-AMPKα1-DN loaded neuronal-targeted dendritic cell-derived sEVs.
a, Western blotting using antibodies against Lamp2b in native and Lamp2b-RVG sEVs. b, Quantification of Lamp2b levels in native (n = 4 samples) and Lamp2b-RVG (n = 5 samples) sEVs in % of native control; P = 0.00031. c, Western blotting using antibodies against GRP94 in Jaws II cells (lane 1), unmodified native sEVs (lane 2) and Lamp2b-RVG sEVs (lane 3). d, Circular representation of the SF1-AMPKα1-DN encoding plasmid. e, Example of curve obtained by nanoparticle tracking analysis of a sample of native (left panel) and SF1-AMPKα1-DN loaded Lamp2b-RVG sEVs (right panel). The graph represents concentration of sEVs (particles/mL) according to the size (nm). f, Electron microscopy image of SF1-AMPKα1-DN loaded Lamp2b-RVG sEVs showing specific round shape and average size of ~70 nm vesicles. g, Agarose gel electrophoresis of native (lane 1), Lamp2b-RVG (lane 2), SF1-AMPKα1-DN loaded Lamp2b-RVG sEVs (lane 3) and negative control H2O (lane 4) of AMPK and GAPDH. h, Agarose gel electrophoresis of SF1-AMPKα1-DN loaded Lamp2b-RVG sEVs treated with DNAse (lane 1), DNAse + Triton X-100 0.2% (lane 2) and Triton X-100 0.2% (lane 3) of AMPK and GAPDH. i, Quantification of pACCα/ACCα in primary astrocytes treated for 24 h with native and Lamp2b-RVG (n = 6 samples/group) sEVs. j, Quantification of pACCα/ACCα in Neuro2A cells treated for 24 h with native and Lamp2b-RVG (n = 3 samples per group) sEVs. Data expressed as mean ± SEM. ***P < 0.001 vs. Control. Statistical significance was assessed by two-sided Student’s t-test.
