Table 1.
Description of selected candidate reference genes and genes of interest.
Table 2.
Description of gene-specific real-time PCR assays.
Figure 1.
Range of quantification cycle (Cq) values of the candidate reference genes.
Boxplot of quantification cycles (Cq) values for each reference gene for mRNA and miRNA normalization in all liver (dark grey box) and small intestine (light grey box) samples was assessed (n = 12). The box indicates the 25% and 75% percentiles. Whiskers represent the maximum and minimum values. The median is depicted by the line across the box.
Figure 2.
geNorm analysis of candidate reference genes for mRNA normalization.
Genes presented on the x-axis in the order of increasing stability (M value on y-axis) for liver and small intestine (A, B). Determination of the optimal number of control genes for normalization in liver and small intestine (C, D).
Table 3.
Ranking of the candidate mRNA RGs according to their stability value using geNorm, NormFinder, and BestKeeper algorithms.
Figure 3.
geNorm analysis of candidate reference genes for miRNA normalization.
Genes presented on the x-axis in the order of increasing stability (M value on y-axis) for liver and small intestine (A, B). Determination of the optimal number of control genes for normalization in liver and small intestine (C, D).
Table 4.
Ranking of the candidate miRNA RGs according to their stability value using geNorm, NormFinder, and BestKeeper algorithms.
Figure 4.
Effects of different normalization approaches on the expression of NQO1.
Fold expression changes of NQO1 gene in liver after green tea catechins treatment was normalized to individual or combined reference genes. Error bars show the standard error calculated from three biological replicates. Stars indicate the significant (P<0.05) difference identified by uncorrected Fisher’s LSD test in multiple comparisons after two-way ANOVA.
Figure 5.
Effects of different normalization approaches on the expression of miR-221 and miR-29b.
Fold expression changes of miR-221 (dark grey columns) and miR-29b (light grey columns) in MSG-obese mice were normalized to individual or combined reference genes. Error bars show the standard error calculated from three biological replicates. Stars indicate the significant (P<0.05) difference identified by uncorrected Fisher’s LSD test in multiple comparisons after two-way ANOVA.