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During the seven days, the new lung area had been flushed having cool PBS to remove bloodstream and you will thumb suspended within the drinking water nitrogen. 5-15 milligrams out of flash-suspended lung cells try incubated which have 0.2 mM CMH inside KHB which includes 100 μM inside two hundred μL away from overall frequency for one h during the 37° C. Supernatant is actually accumulated and you will listed in PTFE tube and you will work at during the 77 K on the digit dewar. (A) Nitroxide amount (μM out of nitroxide normalized to one milligrams away from cells). (B) Associate spectra away from nitroxide within the lung tissues from PBS- and you can Bleo-managed rats. (A) Raw 264.7 muscle sparked that have ten μM PMA and you may EPR spin probe, CMH 0.25 mM (50 minute at the 37 °C) which have (black) otherwise instead of (red) pretreatment having 30 You/mL SOD.
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