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Fig. 2 | Journal of Biomedical Science

Fig. 2

From: Age-associated reduction of cell spreading induces mitochondrial DNA common deletion by oxidative stress in human skin dermal fibroblasts: implication for human skin connective tissue aging

Fig. 2

Reduced cell spreading induces mtDNA common deletion associated with increased reactive oxygen species (ROS) in human skin dermal fibroblasts. a Representative images of dermal fibroblasts treated with Lat-A (see Methods for details), which impairs actin polymerization and fibroblast spreading resulting in a rounded cell shape. Dermal fibroblasts were stained with phalloidin and were imaged by fluorescence microscopy. Red fluorescence delineates cell cytoplasm; blue fluorescence delineates nuclei. Bars = 50 μm. b The relative cell surface areas were quantified by ImageJ. Mean ± SEM, N = 4, *p < 0.05. c Reduced cell spreading induces mtDNA common deletion. mtDNA common deletion was determined by real-time QPCR (see details for Methods). Mean ± SEM. N = 4 *p < 0.05. d Cells were stained with MitoTracker fluorescent dye and were imaged by fluorescence microscopy. Red fluorescence delineates gross morphology of mitochondria; blue fluorescence delineates nuclei. Bars = 50 μm. e and f Reduced cell spreading induces ROS. Intracellular ROS levels were measured by Redox Sensor Red fluorescence and quantified by ImageJ (see Methods for details). Red fluorescence indicates ROS and blue fluorescence delineates nuclei. Mean ± SEM. N = 4, *p < 0.05. g Reduced cell spreading induces mitochondrial ROS. Cells were double stained with Mitotracker (green) and RedoxSensor (red); blue fluorescence delineates nuclei. Bars = 100 μm

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