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The cycle threshold (CT) for the target genes ( SIRT-1 and CYP2E1 ) and the endogenous control GAPDH for each sample were detected and the relative quantitation of SIRT-1 and CYP2E1 mRNA expression levels was estimated by normalizing CT of each gene with the CT of GAPDH in each sample using the 2 CT method ( Hematoxylin and eosin stain (H& E) This stain was used for evaluation of liver architecture, inflammatory cell infiltration, vascular congestion and hepatocyte necrosis, apoptosis, steatosis, vacuolation, edema and malignant changes ( Perls prussian blue stain This stain was used for detection of iron in hepatic tissue

Two contrasting sources here and here