Fig. 5 | Signal Transduction and Targeted Therapy

Fig. 5

From: Bone-targeting engineered small extracellular vesicles carrying anti-miR-6359-CGGGAGC prevent valproic acid-induced bone loss

Fig. 5

Construction of E-sEVs. a Schematic diagram showing the construction process of E-sEVs. b qRT-PCR analysis of Lamp-2b and CX3CL1 as well as anti-miR-6359 expression levels in osteoclast precursors infected with the Lamp-2b and CX3CL1-overexpressed lentivirus or negative controls (n = 4). c Western blot analysis of the expression of Lamp-2b and CX3CL1 proteins in sEVs derived from ECs after infection. d qRT-PCR analysis of anti-miR-6359 in sEVs (n = 4). e, f TRAP staining of osteoclasts (e), and their corresponding quantifications (f) (n = 3). Scar bar: 100 μm. g, h qRT-PCR detection of SIRT3 (g), Nfatc1, Ctsk and Trap (h) mRNA levels in osteoclast precursors following corresponding treatments (n = 4 or 6). i qRT-PCR analysis of Lamp-2b and CX3CL1 as well as anti-miR-6359-CGGGAGC expression levels in osteoclast precursors infected with pLenti-U6-anti-miR-6359-CGGGAGC-EF1N promoter-Lamp-2b (CX3CL1-Extra-sv40-puro)-sv40-puro lentivirus or negative controls (n = 4). j Western blot analysis showing the Lamp-2b and CX3CL1 protein levels in sEVs. k qRT-PCR analysis of anti-miR-6359-CGGGAGC in sEVs (n = 4). l SEVs morphology. Scar bar: 100 nm. m SEVs diameter distribution. n SEVs marker analysis by western blot. (o) Organ fluorescence of mice injected intravenously with PBS, dissolved DiR equivalents, DiR-labeled sEVs and DiR-labeled E-sEVs. p The femurs from mice treated with DiR-labeled E-sEVs were harvested and subjected to immunofluorescence staining for LY6G, CD11b, CD3, CD90 and CX3CR1. Scar bar: 5 μm. Data are presented as the mean ± SEM (n ≥ 3) (*p < 0.05; **p < 0.01; ***p < 0.001)

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