Fig. 3
TPFPD-related chronic-type inflammatory stimulation of microglia is associated with lower cytokine but increased glutamate release. A Representative images of CD11b immunostaining (grey) and Hoechst nuclear staining (blue) of microglial cells following inflammatory stimulation by PD patient-derived αSYN fibrils (1.5 µM; FPD) or TPFPD as compared to nonstimulated cells (NSC). Note that FPD and TPFPD induce similar morphological changes of microglial cells. Scale bar: 50 µm. B Quantification of TNFα, IL-6, IL-10 and IL-1β release by microglial cells exposed or not (NSC) to TNFα (17 ng/mL), PGE2 (1 µg/mL), TNFα + PGE2 (TP), FPD (1.5 µM), TNFα + PGE2 + FPD (TPFPD) or LPS (10 ng/mL). The data are represented as the means ± SEM (n = 3–6). *p < 0.05; **p < 0.01; ***p < 0.001 (Tukey’s test). C Quantification of ROS production measured by the NBT reaction in microglial cells exposed or not (NSC) to FPD (1.5 µM), TPFPD or LPS (10 ng/mL). The data are expressed as a % of the NSC control. The bars are the means ± SEM (n = 6). *p < 0.05 vs. NSC (Student’s t-test). D Quantification of glutamate release by microglial cells exposed or not (NSC) to FPD (1.5 µM), TPFPD or LPS (10 ng/mL). The data are expressed as a % of the NSC control. The bars are the means ± SEM (n = 3–6). *p < 0.05 vs. TPFPD (Tukey’s test)