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作者应用ELISA、Western blot、FCM实验技术，通过实验分析得出：EVs参与CD4+ T手机介导的B手机应答。

Figure 1. Inhibition of EVs release impairs the function of CD4T cells in vitro. A) Bicinchoninic acid (BCA) protein assay of total protein (top) and immunoblot analysis of CD63 (bottom) in purified EVs from supernatants of CD4T cells treated with a control vehicle (dimethyl sulfoxide (DMSO)) or 10, 20, or 40 ×10−6 m GW4869 for 48 h. B) A schematic of the Transwell coculture model with B cells in the upper chamber and CD4T cells in the lower chamber of the well. A porous (0.4 m) membrane allows the transfer of EVs but precludes direct cell contact. C) Flow cytometry analysis of CD86 and MHCII expression on the surface of B cells. D) Total IgG levels in B cell supernatants were analyzed by enzyme-linked immunosorbent assay (ELISA). E) A schematic of the utilization of anti-CD63 microbeads to remove EVs in CD4T cell supernatant. F) The removal efficiency of CD63-positiveEVs in the supernatant was evaluated by western blotting. G) CD86 and MHCII expression on the surface of B cells cultured with different supernatants for 48 h was analyzed by flow cytometry (FCM). H) B cells were cultured with different supernatants for 72 h, and the total IgG in the supernatant was analyzed by ELISA. *P 0.05, **P 0.01, and ***P 0.001 (Student’s t-test). The data are from three independent experiments (A (top), C, D, F (left),G, and H; mean and s.e.m.) or are representative of three independent experiments (A (bottom) and F (right)).
 

• 作者单位：江苏大学医学院

• 论文期刊名：Advanced Science

• 影响app：15.8

• 应用产品：Mouse IgG ELISA Kit

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