Researchers identify protein that drives the development of alcohol-associated liver disease iumedschool NatureComms
. Briefly, minced tissue was homogenized with a Dounce homogenizer in Buffer 1 and the homogenate was centrifuged at 740 g for 5 min. The supernatant was separated into crude mitochondrial pellet and ER containing supernatant by centrifugation at 9000 g for 10 min. The supernatant was collected for ER isolation, whereas, the crude mitochondrial pellet was resuspended in buffer 2 , and centrifuged at 10,000 g for 10 min.
. The architecture of mitochondria and ER in the EM images were reconstructed graphically using GIMP software , and analyzed using ImageJ .Cells were seeded on 12 mm glass coverslips in 24 well plates. At the end of the experiment, cells were washed with PBS, fixed with 4% paraformaldehyde, permeabilized , and incubated with primary antibodies overnight at 4 °C.
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