Fig. 6 | Signal Transduction and Targeted Therapy

Fig. 6

From: Mesenchymal stem cell-derived small extracellular vesicles mitigate oxidative stress-induced senescence in endothelial cells via regulation of miR-146a/Src

Fig. 6

miR-146a rescue senescent HUVEC by downregulating Src signal pathway. a Phospho-kinase antibody array was performed on protein lysates from HUVEC pretreated with (S + 0 ng/μl MSC-sEV, S + 200 ng/μl MSC-sEV (30 min), S + 200 ng/μl MSC-sEV (4 H)) or without H2O2 (control). b Quantitation of different phosphorylation levels of P38, Akt1/2/3, Erk1/2, Src, JNK1/2/3, c-Jun which calculated from the array pixel density. The chosen protein was highlighted by red boxes in (a). c Western blot analysis showing phosphorylation of P38, AKT1/2/3, Erk1/2, Src, JNK1/2/3, c-Jun. d Western blots analysis showing the change of senescence markers P16, P21, P53, and LMNB1 in HUVEC treated with an inhibitor of JNK1/2/3(SP600125), Src (Saracatinib), P38 (SB203580), AKT1/2/3 (GSK690693), and Erk1/2 (U0126) before treated with H2O2 (50 μM, 2 h). e Western blots analysis showing the phosphorylation change of downstream of Src signal pathway VE-cadherin and Caveolin after co-cultured with MSC-sEV (200 ng/μl) for 30 min and 4 H compared with group control and group S (without MSC-sEV). f Western blots analysis showing the phosphorylation of Src, VE-cadherin, and Caveolin (downstream of Src) in high-glucose-induced senescent HUVEC. HUVEC was co-cultured with MSC-sEV (200 ng/μL, 48 h) after treated with high glucose (30 mM, 48 h). g Representative immunofluorescence staining images of positive cells of p-Src, p-VE-cadherin, p-Caveolin-1 (red staining), and CD31-positive cells (green staining) on paraffin-embedded sections of senescent C57BL/6mice dorsal skin injected PBS (control) or MSC-sEV (scale bar, 100 μm). h Quantitation of positive cells of p-Src, p-VE-cadherin, p-Caveolin-1 in CD31-positive cells. i Representative immunofluorescence staining images of positive cells of p-Src, p-VE-cadherin, p-Caveolin-1 (red staining), and CD31-positive cells (green staining) on paraffin-embedded sections of type-2 diabetes model C57BL/6mice dorsal skin injected PBS (control) or MSC-sEV (scale bar, 200 μm). j Quantitation of positive cells of p-Src, p-VE-cadherin, p-Caveolin-1 in CD31-positive cells. k Western blots analysis showing the phosphorylation change of Src, VE-cadherin, and Caveolin (downstream of Src) after transfected mimic-146a in HUVEC

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