Fig. 3

Rapamycin treatment restores the Δψ_m and decreases the production of ROS in CoCl₂-exposed cells. a Rapamycin treatment prevents hyperpolarization of the mitochondrial membrane. HT22 cells were untreated or treated with 250 µM CoCl₂ with and without 500 nM rapamycin for 24 h. 500 nM TMRM dye was added during the final 30 min of treatment. Cells were washed with PBS and fluorescence was measured using a PHERAstar Microplate Reader with a 590-50/675-50 filter. Relative fluorescence intensities were obtained by subtracting background signal from treated cells without TMRM/DHE and normalizing to the percent of viable cells. ^##p < 0.01 versus control; ^###p < 0.001 versus control; ***p < 0.001 versus CoCl₂ alone. b Rapamycin treatment decreases ROS production. Cells were treated and analyzed as in A except 5 µM DHE dye was added instead of TMRM. ^#p < 0.05 versus control; ^###p < 0.001 versus control; ***p < 0.001 versus CoCl₂ alone