Fig. 4

OL-HSPB8-EVs reduce reactive oxygen species upon PMA stimulation. A–D Flow cytometry monitoring of ROS activity assay in human microglia C20 cell line (A, B) and a primary mixed neural culture (C, D) using the DCFH-DA detection dye. Data are presented as mean fluorescent intensities ± SEM (n = 3 biological replicates per group). Graphpad was used to determine significance by using a one-way ANOVA statistical analysis (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). E Relative qPCR expression of ROS-related genes SOD1 and SOD2 in human C20 cells stimulated with OL-EV or OL-HSPB8-EV either non-activated or TNFα/IL1β activated. As a positive control, the ROS inducer PMA was used. Values are represented as the mean ± SEM of three independent biological replicates. Data were normalized to non-stimulated equals 1. One-way ANOVA was used to determine statistical significance where P < 0.05 was considered statistically significant