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. Author manuscript; available in PMC: 2013 Sep 1.
Published in final edited form as: Free Radic Biol Med. 2012 Jul 8;53(5):1111–1122. doi: 10.1016/j.freeradbiomed.2012.06.042

Fig.5. ER stress induces release of cytochrome c from hRPE mitochondria.

Fig.5

A. hRPE cells treated with 3 μg/ml TM for 24 h and 10 μg/ml TM for 6 h were used for analysis of cytochrome c by immunofluorescence and western blot. The distribution of cytochrome c (green) and Mito Tracker (red) by confocal microscopy showed release of cytochrome c into cytosol under ER stress. B. Western blot analysis of cytochrome c in isolated mitochondrial and cytosolic fractions of hRPE cells exposed to 3 μg/ml TM for 24 h. The blot shows that especially after 24 h TM treatment, a significant amount of cytochrome c is released from mitochondria to cytosol (upper panel). The lower panels show the fold changes calculated by normalization of band density with anti-COX IV (mitochondrial protein marker) and anti-Tubulin (cytosolic protein marker), which further confirmed a >50% increase of cytochrome c in cytosol. Values are the mean±S.D.from three independent experiments. * p< 0.05. Bar: 10 μm.