FIGURE 4.

Transfection of siRNA-MYH9, but not siRNA-MYH10 or siRNA-MYH14 inhibited exosome release from microglial cells stimulated by LPS. For transfection, microglial cells were sub-cultured in 24-well plates at a density of 3 × 10⁵ cells/ml. The extracted exosomes were grouped based on transfection with siRNA or siRNA into microglial cells. Whether control or LPS stimulation cells were transfected with siRNA-MYH9, siRNA-MYH10, or siRNA-MYH14 at a final concentration of 100 nM for 48 h. Microglial cells stimulated by 1 mg/ml LPS for 24 h, microglial cells transfected with siRNA followed by 24 h LPS stimulation, microglial cells transfected with siRNA negative control followed by 24 h LPS stimulation. (A) Nanoparticle tracking analysis for the concentration of exosomes. (B) Flow cytometry analysis for CD9-labeled positive rate, CD81-labeled positive rate, or control FITC-IgG-labeled positive rate. Nine independent experiments were analyzed. A histogram depicted the quantitative representations of the protein expression levels of CD9 and CD81 for each group. The data were averages with SD, n = 9. ^** p < 0.01 vs. control microglial cells; ^## p < 0.01 vs. microglial cells stimulated with LPS.