Fig. 4
Effect of tigecycline/tetracycline treatment on mitochondrial function. Cells were treated with Tig/Tet for 24 and 48 h, and equal numbers of cells (10,000 each in triplicates) were used for analysis of mitochondrial parameters along with untreated controls. A Total ROS levels were quantified by staining of cells with 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA), and B mitochondrial superoxide levels were measured by MitoSOX™ staining, followed by fluorescence plate reading. For both measurements, the cells were counterstain with nuclear stain Hoechst33342 for normalization. C Changes in mitochondrial membrane potential were measured by JC1 staining and quantifying the J aggregates red fluorescence at 590nm and normalized with nuclear stain Hoechst33342 readings in a plate reader. D Change in total cellular ATP content was measured in cells with a luciferase-based one-step ATP detection through luminescence and normalized with equal cell count. Relative levels of different parameters are shown individually in graphs for a Tig and b Tet. *P<0.05**P<0.01 and ***P<0.001 vs. Untreated. Abbreviations: ROS, reactive oxygen species; ATP, adenosine triphosphate; JC1, 5,5,6,6′-tetrachloro-1,1,3,3′-tetraethylbenzimi-dazoylcarbocyanine iodide