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Fig. 2 | Journal of Translational Medicine

Fig. 2

From: Renal temperature reduction progressively favors mitochondrial ROS production over respiration in hypothermic kidney preservation

Fig. 2

Temperature effects on HEK293 cells. a State three oxygen consumption at normothermic (37 °C) and hypothermic (4 °C) circumstances in coupled and uncoupled HEK293 cells. Based on three independent experiments, expressed as rel. to normothermic, error bars represent SEM. #p < 0.001 (Students T test). b Lipid peroxidation (MDA) at different temperatures in HEK293 cells. N = 6, expressed as MDA levels rel. to normothermic, error bars represent SEM. #p < 0.001 compared to 37 °C (ANOVA with Bonferroni posthoc). c Mitochondrial membrane potential in normothermic, hypothermic and rewarmed (rew) HEK293 cells. N = 6, expressed as JC1 ratio RFU rel. to 37 °C (red [590 nm]/green [529 nm]), error bars represent SEM. #p < 0.001 compared to 37 °C (ANOVA with Bonferroni posthoc). FCCP (carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone 1 μM) as uncoupled control. d Mitochondrial superoxide production in HEK293 cells, incubated for 90 min at 37°, 22° or 4°. N = 6, expressed as mitosox RFU, error bars represent SEM. #p < 0.001 compared to 37 °C (ANOVA with Bonferroni posthoc). e Lipid peroxidation in H2O2 stimulated normothermic and hypothermic treated HEK293 cells. N = 6, expressed as MDA levels rel. to normothermic, error bars represent SEM. *p < 0.05 compared to control (Student t test). f HEK293 survival after 3 h H2O2 exposure at different concentrations and temperatures. N = 3, expressed as Neutral Red absorbance rel. to untreated control. *p < 0.05 compared to non-treated (Student t test), #p < 0.001 (Students T test). g MnSOD protein expression in normothermic and 6 h hypothermic (4 °C) HEK293 cells. Expressed as corrected values for actin, rel. to 37 °C, error bars represent SEM. *p < 0.05 (Student t test). Full membranes shown in Additional file 1: Figure S3B. h MDA levels in trolox treated 6 h hypothermic HEK293 cells. N = 3, expressed as MDA levels rel. to normothermic, error bars represent SEM. #p < 0.001 compared to 37 °C (ANOVA with Bonferroni posthoc)

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