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 H
2O
2 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 H
2O
2 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)