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. Author manuscript; available in PMC: 2007 Aug 14.
Published in final edited form as: J Biol Chem. 2002 Sep 5;277(45):43377–43388. doi: 10.1074/jbc.M206373200

Fig. 4. Quantification of JC-1 accumulation in mitochondria (red fluorescence) and cytoplasm (green fluorescence) in cisplatin-treated RPTC.

Fig. 4

RPTC monolayers were exposed to 50 μm cisplatin for 24 h and loaded with 10 μm JC-1 for 30 min at 37 °C. After loading, media were aspirated and monolayers kept on ice, washed twice with ice-cold PBS, scraped off culture dishes, washed, and resuspended in PBS. Fluorescence was analyzed by flow cytometry (BD Biosciences FACSCalibur) using excitation by 488 nm argon-ion laser. The JC-1 monomer (green fluorescence) and J-aggregate (red fluorescence) were detected in FL1 (emission, 525 nm) and FL2 (emission, 590 nm) channels, respectively. A, the effect of cisplatin on JC-1 accumulation in RPTC mitochondria. B, the effect of inhibition of ERK1/2 (50 μm PD98059) on cisplatin-induced accumulation of JC-1 in mitochondria. C, the effect of inhibition of PKC-α (10 nm Go6976) on cisplatin-induced accumulation of JC-1 in mitochondria. Experiments were performed five times with comparable results.