a, We repeated the experiment in Fig. 2c with n=4 independent primary
cultures per group. Similar results were obtained. The extracellular
mitochondria-depleted astrocyte media (mdACM) group was significantly
different compared to the ACM group. Furthermore, in this repeated
experiment, there was also statistical significance between controls
(OGD-damaged neurons alone) versus those treated with
mitochondria-containing astrocyte media (ACM), and there was no
statistically significant worsening when comparing control versus
mitochondria-depleted groups (mdACM). Taken together, these two separate
experiments suggest a modest but statistically significant neuroprotection
induced by astrocyte-derived mitochondria. All values are mean
+/− SEM. b, Mitotracker Red CMXRos (200 nM) was
incubated without astrocytes to obtain no-cell-derived media (negative
control). Media was collected and further incubated with neurons following
oxygen-glucose deprivation. After 24 hours, there was no mitochondrial
signal observed. Scale: 100 μm.