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Fig. S1. Representative traces of mitochondrial Ca2+ responses following stimulation of adult WT and CyPD-KO cortical neurons with DHPG, and quantification of mitochondrial Ca2+ levels prior to stimulation and at peak in response to DHPG.

Fig. S2. Pretreatment with the mitochondrial uncoupler FCCP (450 nM) for 3 min abolishes mitochondrial Ca2+ uptake following perfusion with ATP in neurons prepared from WT and CyPD-KO, absence of mitochondrial Ca2+ responses following depolarization in the absence of extracellular Ca2+, and pretreatment with mitochondrial uncoupler FCCP (450 nM) for 3 min abolishes mitochondrial Ca2+ uptake in neurons prepared from either genotype following depolarization with KCl.

Fig. S3. Mitochondrial morphology in neurons remains unaltered following perfusion with ATP and KCl.

Fig. S4. Mitochondrial Ca2+ responses following simultaneous stimulation with either ATP or depolarization in WT and CyPD-KO adult cortical neurons treated with CsA.

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