Relative quantification of iNOS, COX-2, aggrecan and collagen type II signals was accomplished by normalizing each target gene to GAPDH and to the calibrator sample by a comparative Ct approach [ 47, 48].
For each of the biological samples, gene expression was quantified by normalizing each target gene against the expression of the reference gene, GAPDH, using the Q-GENE statistical analysis package [ 52].
Relative quantification of MMP-3, MMP-13, collagen type II and fibronectin signals was accomplished by normalizing each target to the reference gene, GAPDH and to the calibrator sample (unstrained, untreated sample) by a comparative Ct approach [ 29].
Relative quantification of collagen type II, aggrecan, COX-2 and iNOS signals were estimated by normalizing each target to the reference gene, GAPDH, and to the calibrator sample (untreated, unstrained condition) by a comparative Ct approach.
Relative quantification of iNOS, COX-2, aggrecan, and collagen type II signals were accomplished by normalizing each target to the reference gene, GAPDH, and to the calibrator sample by a comparative Ct approach.
Relative quantification of Npr2, Npr3, CNP, aggrecan and collagen type II signals were estimated by normalizing each target to the reference gene, GAPDH, and to the calibrator sample (unstrained, untreated control) by a comparative Ct approach.
Relative expression levels for each gene of interest were calculated by normalizing the target gene transcript level (Ct) to the respective GAPDH level as described previously (2−ΔΔCt formula, Perkin Elmer User Bulletin #2).
The relative amount of target mRNA was determined using the comparative threshold (Ct) method by normalizing target mRNA Ct values to those for β-Actin (Δ Ct).
Relative expression levels of samples were calculated by normalizing target levels to the endogenously expressed housekeeping gene (beta-actin).
The relative transcript levels were calculated by normalizing target gene expression to control gene expression, where expression of the control gene was set to 1 [ 62, 63].
The relative expression levels were calculated by normalizing the targets to the endogenously expressed housekeeping gene (β-actin).
