The data are summarized in Figure 2a — c. Values for the housekeeping
gene cyclophilin A were highly consistent across all cells analyzed.
Towards the center of the colony, ∼ 40 % of the cells had lost expression of all pluripotency markers, and we found a number of cells that were null - expressing no markers of pluripotency or lineage commitment, only the housekeeping
gene cyclophilin a. However, many cells in this region still expressed Oct - 4.
Not exact matches
For expression analysis in mice, we used microarray data as described above to select two internal control
genes,
cyclophilin B (Cphn2) and ribosomal protein S3 (Rps3).
Q - PCR showed that ULK1 mRNA levels were increased 3 -7-fold in DAXX K / D samples, using the
cyclophilin (CPH) internal control
gene for normalization (Figure 6C).
Data are normalized to
cyclophilin and expressed as ΔCt (marker
gene —
cyclophilin).
Expression of four stem cell marker
genes normalized to
cyclophilin in nine single cell equivalents prepared from a pool of nine lysed HES3 cells.
CT values for
cyclophilin were somewhat more variable in cells isolated by flow cytometry and frozen prior to analysis, compared to freshly isolated and unmanipulated cells, but the gradients in patterns of
gene expression were similar for cells isolated by the two techniques (Figure 3a — d).
For each sample, expression of marker
genes was normalized to
cyclophilin.