Fig 3.

B355252 reduces both production of ROS and restores mitochondrial membrane potential after CoCl₂-induced hypoxia A) B355252 decreases the elevated ROS production observed during CoCl₂-induced hypoxia. The presence of ROS was determined using CellROX Deep Red Reagent. HT22 cells treated for 24h with 300 μM CoCl₂ and 2.5 μM B355252 (2h prior) before adding 5 µM CellRox Deep Red Reagent and incubating at 37°C for 30 min. B) Mitochondrial membrane potential is restored after B355252 treatment. Quantification of the mitochondrial membrane potential was done using Tetramethylrhodamine, methyl ester (TMRM) in HT22 cells. Cells were treated with CoCl₂ and B355252 as in A before adding 500 nM TMRM and incubating cells at 37^oC for 30 min. In both A and B, fluorescence was measured using a PHERAstar Microplate Reader with a 590-50/675-50 filter. The results of both the CellRox and TMRM fluorescence measurements were normalized against the cell viability to obtain the relative fluorescence values with control cells set at 100% fluorescence. One-way ANOVA followed by Bonferroni's post-test was used for analysis. ### p<0.001 vs. control. ** p<0.01 vs CoCl₂ alone. *** p<0.001 vs. CoCl₂ alone.