Relative transcript abundance was assessed by normalizing the cycle threshold (Ct) of the MMP gene of interest with that of the endogenous control, β-Actin.
Relative expression levels were determined by normalizing the cycle threshold (Ct) value of each target gene by the mean Ct value of three reference genes, i.e., glyceraldehyde 3-phosphate dehydrogenase (GAPDH), ribosomal protein S19 (RPS19), and TATA-binding protein (TBP).
The quantity of mRNA was calculated by normalizing the cycle threshold (CT) value of MIF to the CT of the housekeeping gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), in the same sample according to the following formula: the average GAPDH CT was subtracted from the average MIF CT; the result represents the ΔCT.
The quantity of mRNA was calculated by normalizing the Cycle Threshold (CT) value of MIF to the CT of the housekeeping gene GAPDH in the same sample, according to the following formula: The average GAPDH CT was subtracted from the average MIF CT; the result represents the ΔCT.
The ΔCTs for each sample and target were obtained by normalizing the threshold cycle number (CT) to the constitutively expressed gene glyceraldehyde-3 phosphate dehydrogenase (GAPDH, amplification efficiency = 95%).
Gene expression was quantified by normalizing the logarithmic cycle threshold (CT) value (2−CT) to the cDNA content of each sample to obtain the expression 2−CT/cDNA content (ng/ml).
Analysis of data was performed by normalizing the threshold cycle number (Δct) of the gene of interest to the Δct for the housekeeping gene, 18S ribosomal RNA, using the 2-ΔΔct method and expression HFC values relative to LJT, MJT, HJT values and expressing in arbitrary units.
The relative mRNA levels were determined by normalizing the PCR threshold cycle number with Actin and 18S RNA.
The relative mRNA levels were determined by normalizing the PCR threshold cycle number of each gene with that of the EF-1α and ACTIN as reference genes, according to GeneEx software (http://www.multid.se/order/bioeps.php;) and the data were the average of three replicates.
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