Figure 8.

TDP-43 up-regulation enhances neuronal vulnerability to death by microglia-mediated cytotoxicity. (A and B) TDP-43 (WT and mutants)–transfected BV-2 cells were stimulated with LPS. 12 h after stimulation, total RNA was extracted with TRIZOL. The total RNA samples were then subjected to real-time quantitative RT-PCR for IL-1β (A) and Nox-2 (B). Error bars represent mean ± SEM from five different experiments. Statistical analysis was performed by two-way ANOVA with Bonferroni adjustment. (C) Primary microglial cells from TDP-43^WT, TDP-43^A315T, TDP-43^G348C, and B6 nontransgenic mice (Ntg) were stimulated or unstimulated with H₂O₂. Immunoblots were run to determine the levels of various proteins using specific antibodies as indicated. A representative blot from two independent experiments is shown. (D–F) Primary cortical neurons from TDP-43^WT, TDP-43^A315T, TDP-43^G348C, and control B6 nontransgenic mice were incubated with the conditioned media (con. media) derived from primary microglial cells treated with 100 ng/ml LPS. 12 h after challenging cortical cells, cell culture supernatants were used for LDH assay (D). ROS production was determined by H2DCFDA fluorescence (E), and nitrite production was evaluated by Griess reagent (F). Error bars represent mean ± SEM from four independent experiments.