Figure EV2. Mitochondria are dysfunctional in Ifnb^−/− neuron and brain, but this is not due to alterations of complex II function.

• A
  Quantification of ATP on aging from 6 to 18 days in Ifnb ^+/+ and Ifnb ^–/– CNs. Error bars are mean + SEM from 3 independent experiments.
• B
  Monolayer of mitochondria after isolation, labeled with MitoTracker Green and TMRE, showing intact and polarized mitochondria.
• C, D
  Respiration of isolated mitochondria from Ifnb ^+/+ and Ifnb ^–/– whole brains. OCR measurements under Complex II substrate (succinate supplemented with rotenone to inhibit Complex 1 activity) were obtained at baseline (B) and on addition of ADP, oligomycin, FCCP, and rotenone/antimycin A to capture S3, S4o, S3u, and nonmitochondrial respiration. N = 3–4 mice per group. Bar graphs show mean + SEM.
• E
  ROS levels based on mean fluorescence intensity of DCFDA in DIV6 Ifnb ^+/+ and Ifnb ^–/– CNs.
• F
  Immunofluorescence of 8OHdG in DIV6 CN cultures. Neurons were labeled with Nissl. Scale bars equal 50 μm.
• G
  Expression of optineurin mRNA in DIV6 Ifnb ^+/+ and Ifnb ^–/– CNs, quantified by qPCR.
• H
  Expression of optineurin mRNA in Ifnb ^+/+ and Ifnb ^–/– brainstems, quantified by qPCR.

Data information: For all graphs, error bars mean SEM; *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 by t‐test (A; G; H) or two‐way ANOVA (E) with t‐test as post‐hoc.