Fig 1: Exosomal miR-21 derived from hypoxic ADSCs activates PI3K/AKT signalling by targeting SPRY1 in the HUVECs. A Schematic representation of the putative binding sites for miR-21-5p in the SPRY1 3′-UTR. Predicted consequential pairings of the target regions and miR-21 (underlined) were based on data from TargetScan (www.targetscan.org). B Luciferase reporters containing wild-type or mutant 3′-UTR of SPRY1 genes were co-transfected with miR-21-NC or miR-21 mimics into HEK293 cells. Two days after transfection, the dual luciferase activity was measured. C–G qRT-PCR was performed to measure the mRNA level of SPRY1, PI3K, AKT VEGFA and HIF-1α in the HUVECs in different groups. H Western blot was performed to measure the level of SPRY1, PI3K, AKT VEGFA and HIF-1α in the HUVECs in different groups
Fig 2: GleMA loaded with hypo-ADSC-Exos enhanced local microvascular network formation and osteoporotic fracture healing in vivo via targeting SPRY1. A Schematic illustration of the process of in vivo treatment using hypoxia-pretreated ADSC-derived exosomes. B SEM images of GelMA loaded with different exosomes. C, D Degration and protein released ratio of GelMA loaded with exosomes. E, F Representative X-Ray images and morphometric analysis of osteoporotic fractures after hypo-ADSC-Exo treatment on 4 week and 8 week. G, H Micro-CT images of osteoporotic fractures after hypo-ADSC-Exo treatment and morphometric analysis of new bone volume (BV), tissue volume (TV), and bone mineral density (BMD). n = 6. I Haematoxylin and eosin (H&E) staining on the bone fracture region after PBS, ADSC-Exo and hypo-ADSC-Exo treatment. n = 6. J, N Micro-CT 3D reconstruction images of angiographic images of fracture area after hypo-ADSC-Exo treatment. Scale bar, 20 μm. K, L Immunofluorescence staining and quantitative analysis results showed the CD31 and EMCN staining on the bone fracture region. Scale bar, 50 μm. n = 6. M The serum levels of VEGFA, OCN, BALP and CTX-1. n = 6. O SPRY1 immunohistochemistry staining images and quantitative analysis on the bone fracture region after PBS, ADSC-Exo and hypo-ADSC-Exo treatment. Scale bar, 50 μm. n = 6. P VEGFA immunohistochemistry staining images and quantitative analysis on the bone fracture region after PBS, ADSC-Exo and hypo-ADSC-Exo treatment. Scale bar, 50 μm. n = 6. (Data are presented as the means ± SD; *p < 0.05; **p < 0.01; ***p < 0.001)
Supplier Page from Abcam for Mouse VEGF ELISA Kit