Table 1.
Descriptions of the fourteen candidate reference genes studied.
Figure 1.
Characteristic cellular morphology and marker gene expression for ES, TS and XEN Stem Cells.
a–c Light micrographs of representative ES (a) TS (b) and XEN (c) stem cell lines used in this study. d) Expression of transcription factors characteristic of each of the stem cell lineages.
Table 2.
Candidate reference genes ranked in order of their stability.
Figure 2.
Relative expression of the fourteen candidate genes between all three genetic backgrounds of ES, TS and XEN stem cells.
CT values for each measured transcript were normalized to the geometric mean of Pgk1, Sdha and Tbp, and then graphed as relative values using methods described [13], [34], [35]. Error bars represent the standard error of the mean.
Table 3.
Consensus of the stability rankings for pair-wise comparisons between the stem cell types.
Figure 3.
A. Relative expression of the fourteen candidate genes throughout differentiation of all three genetic backgrounds of ES cells examined.
CT values for each transcript were measure on Day 0, Day 4 (embryoid body) and Day 8 and were then normalized to the geometric mean of Sdha Tbp and Ywhaz. Relative values were determined using methods described previously [13], [34], [35] and graphed. Error bars represent the standard error of the mean. B. Increased expression of fibroblast specific protein 1 throughout ES cell differentiation. Error bars represent the standard error of the mean for three independent replicates.
Figure 4.
Relative expression of the fourteen candidate genes throughout differentiation of all three genetic backgrounds of TS cells.
CT values for each transcript were measured on Day 0 and Day 8 and were then normalized to the geometric mean of Ywhaz, Pgk1 and Hk2. Relative values were determined using methods described previously [13], [34], [35] and graphed. Error bars represent the standard error of the mean. Note that the top third of the graph is in an exponential scale.
Table 4.
Candidate reference genes ranked in order of their stability throughout ES cell differentiation using the NormFinder, GENorm and BestKeepr software tools.
Table 5.
Candidate reference genes ranked in order of their stability throughout TS cell differentiation using the NormFinder, GENorm and BestKeepr software tools.
Table 6.
Description and sequences of the primers used in the in the analysis of both the candidate reference genes and lineage specific transcription factors.