Figure 7.

Inhibiting the UPR^mt abolishes the protective effects of Sphk1 overexpression on neurons and their mitochondria. N2a cells were transfected with Ad-Ctrl or Ad-Sphk1, and then were subjected to OGD/R to simulate cerebral I/R injury in vitro. A. Cell viability was determined with an MTT assay. B. An LDH release assay was used to measure LDH levels in media from N2a cells. C, D. JC-1 probes were used to analyze the mitochondrial membrane potential in N2a cells after OGD/R injury. The red-to-green fluorescence ratio was used to quantify the mitochondrial membrane potential. E. Immunofluorescence was used to measure the production of mitochondrial ROS. F, G. The activity levels of caspase-3 and caspase-9 were determined using ELISAs in N2a cells. *p<0.05 vs. Ad-Ctrl group, #p<0.05 vs. OGD/R+Ad-Ctrl group, @p<0.05 vs. OGD/R+Ad-Sphk1 group.