Table 1.
Primers and relative information of the reference and target genes.
Table 2.
Sequencing results of genes using BLASTN from NCBI against nucleotide collection.
Fig 1.
Distribution of the CT values of the reference genes in the duodenum (A), jejunum (B), ileum (C), colon (D) and whole gastrointestinal tract (E) of piglets. The boxes encompass the 25th to the 75th percentiles. The middle line in a box marks the median. Whisker caps indicate the minimum and maximum values.
Fig 2.
Distribution of the CT values of the reference genes at different growth stages of the gastrointestinal tract.
The boxes encompass the 25th to the 75th percentiles. The middle line in a box marks the median. Whisker caps indicate the minimum and maximum values.
Table 3.
A comparison of CT values of the 18 reference genes at different GITs and growth stages.
Fig 3.
Gene expression stability and ranking of the reference genes in the duodenum as calculated by GeNorm.
A) Rank order of gene expression stability based on average expression stability values (M) for the reference genes from least stable (left) to most stable (right). B) Pairwise variation analysis (V) to determine the optimal number of reference genes for RT-qPCR data normalization.
Fig 4.
Gene expression stability and rankings of the reference genes in the jejunum as calculated by GeNorm.
A) Rank order of gene expression stability based on average expression stability values (M) for the reference genes from least stable (left) to most stable (right). B) Pairwise variation analysis (V) to determine the optimal number of reference genes for RT-qPCR data normalization.
Fig 5.
Gene expression stability and rankings of the reference genes in the ileum as calculated by GeNorm.
A) Rank order of gene expression stability based on average expression stability values (M) for the reference genes from least stable (left) to most stable (right). B) Pairwise variation analysis (V) to determine the optimal number of reference genes for RT-qPCR data normalization.
Fig 6.
Gene expression stability and rankings of the reference genes in the colon as calculated by GeNorm.
A) Rank order of gene expression stability based on average expression stability values (M) for the reference genes from least stable (left) to most stable (right). B) Pairwise variation analysis (V) to determine the optimal number of reference genes for RT-qPCR data normalization.
Fig 7.
Gene expression stability and rankings of the reference genes in the whole gastrointestinal tract as calculated by GeNorm.
A) Rank order of gene expression stability based on average expression stability values (M) for the reference genes from least stable (left) to most stable (right). B) Pairwise variation analysis (V) to determine the optimal number of reference genes for RT-qPCR data normalization.
Table 4.
Reference genes stability calculated by BestKeeper based on the CT.
Fig 8.
Effect of normalization on ALP expression values in the duodenum (A), jejunum (B), ileum (C), colon (D) and the whole gastrointestinal tract (E) of piglets. The expression of ALP was normalized to 18S, B-actin, HMBS, B2M and HPRT1 and the geometric mean of B2M/HMBS/HPRT1 expression and is shown relative to its expression at different post-weaning stages. Error bars indicate mean ± standard error of the mean (SEM).