Fig. 4.

In vitro production of H₂O₂ and NO by cytokine-activated MNs (a), astrocytes (b), microglia (c) and endothelial cells (d). Cells were isolated from mice at week 18 of progression and plated immediately after. Isolated cells were incubated x 24 ​h before addition of the cytokines. The concentrations used for each cytokine were selected based on the results obtained in Fig. 3. All cytokines were recombinant murine (rmTNFα and rmIFNγ were obtained from Merck; and rmIL1β from Sino Biological Europe GmbH, Eschborn, Germany). H₂O₂ and NOx were measured 6 ​h after adding the cytokines to the culture flasks. Data represent the total amount of NOx and H₂O₂ that accumulated in the medium during the indicated 6 ​h-period. rmTNFα concentrations in the culture medium were 1 (cells isolated from WT mice) or 25 ​pg/ml (cells isolated from SOD1^G93A or FUS^R521C mice). rmIFNγ concentrations in the culture medium were 2 (cells isolated from WT mice) or 100 ​pg/mL (cells isolated from SOD1^G93A or FUS^R521C mice). rmIL1β concentrations in the culture medium were 0.5 (cells isolated from WT mice) or 50 ​pg/ml (cells isolated from SOD1^G93A or FUS^R521C mice). Values are means ​± ​SD of five to six different experiments. ∗P ​< ​0.05 (t-test) comparing cells isolated from SOD1^G93A or FUS^R521C mice versus cells isolated from WT mice.