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Fig. 6. CAP1 knockdown reduces cytochrome c release and ROS accumulation. (A) CAP1-knockdown cells have reduced cytochrome c release when apoptosis is induced. HeLa stable cell lines harboring vector (vector) and CAP1 shRNA (S3-2) treated with STS (1 µM). After 4 hours of treatment, mitochondria were isolated by sucrose gradient centrifugation and subjected to western blots for CAP1 and cytochrome c. VDAC was monitored to normalize total protein. (B) Effect of CAP1 knockdown on ROS accumulation. HeLa wild-type cells, stable cell lines with scrambled S2 or CAP1 shRNA were cultured overnight and treated with 1 µM STS for 1 hour, cells were washed and stained with fresh 10 µM H2DCF-DA (Ros; Invitrogen) for 15 minutes and then examined under an inverted fluorescence microscope. All images were taken with the same exposure time.
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