Article Figures & Data
Figures
- Figure 1.
ROS production in mouse renal proximal tubule cells after treatment with H2O2 in the presence or absence of F7. TKPTS cells were loaded with the oxidant-sensitive fluorescent dye DCFDA as described in Materials and Methods and pretreated with 50 μl/ml F7 for 30 minutes prior to treatment with 400 μM H2O2. Untreated, F7- and 400 μM H2O2-treated controls were also included. ROS production was determined as an increase in DCFDA fluorescence as measured in a fluorescence plate reader at 485nmexc/530nmem. All experiments were performed in triplicate and results expressed as mean±S.D.
- Figure 2.
Mitochondrial depolarization in renal proximal tubule cells after treatment with H2O2 in the presence or absence of F7. TKPTS cells were loaded with the mitochondrial polarity-sensing dye JC-1 (see Materials and Methods) and treated with 50 μg/ml F7 for 30 minutes then 400 μM H2O2 was added. Untreated, F7- and H2O2-treated cells were also processed. Mitochondrial depolarization was calculated as decrease in J aggregate fluorescence/5×105 cells/10 minutes and expressed as a percentage of that of untreated cells. N=3, mean±S.D., *p<0.05 compared to untreated control; #p<0.05 compared to H2O2-treated group.
- Figure 3.
LDH release in renal proximal tubule cells after treatment with H2O2 in the presence or absence of F7. TKPTS cells grown in 12-well-plates were pretreated with 50 μg/ml F7 for 30 minutes then 400 μM H2O2 was added: some wells received F7 or H2O2 treatment only, or were left untreated. After 24 hours' incubation at 37°C, LDH release was determined as described in Materials and Methods. Each experiment was carried out in triplicate: values are expressed as percentage of untreated control (N=3, mean±S.D.) *p<0.05 compared to the untreated control; #p<0.05 compared to H2O2-treated group.
- Figure 4.
Morphology (A) and surviving cell counts (B) of TKPTS cells after treatment with H2O2 in the presence or absence of F7. A, TKPTS cells were pretreated or not with 50 μl/ml F7 for 30 minutes prior to treatment with 400 μM H2O2. After 24 hours' incubation at 37°C, images were taken (magnification: ×100). Images shown are representative of three independent experiments. B, Cell numbers were determined by trypan blue exclusion in cells that were treated in (A). Cell survival was expressed as cell counts of untreated values. *p<0.05 compared to the untreated control; #p<0.05 compared to H2O2-treated group.
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