Figure 5. Mitochondrial electron transport is necessary for the PM-induced acceleration of carotid thrombosis.
(A-F) Mice deficient in Tfam, a nuclear encoded transcription factor necessary for mitochondrial DNA transcription, in alveolar macrophages (CreCD11c/Tfamflox/flox) were compared with control mice. (A) Levels of Tfam mRNA in flow sorted AMs and neutrophils. (B) Mice were treated with PBS or PM (10 μg, intratracheally) and 24 hours later, the time to cessation of carotid artery blood flow after a standardized ferric chloride injury was measured. (C) Wild-type C57Bl/6 mice were treated with the mitochondrially targeted antioxidant MitoTempo (0.7 mg/kg/day, intraperitoneally) 24 hours before and simultaneous with the administration of PM (10 μg/mouse) or PBS and the time to cessation of carotid artery blood flow after standardized ferric chloride injury was measured. (D) Primary alveolar macrophages from the indicated strains of mice were treated with PM and IL-6 levels in the media were measured 24 hours later (n=3, * P<0.05). (E,F) Primary alveolar macrophages from the indicated strains were loaded with MitoSOX and mitochondrial ROS generation was measured continuously after the administration of PM on the stage of an epifluorescent microscope (n= 4, * P <0.05). (G,H) Primary alveolar macrophages were loaded with Fura-2 for measurement of intracellular calcium levels after PM exposure in calcium free followed by calcium replete media. Alveolar macrophages isolated from 3 mice per group (* p<0.05). See also Figure S5.