For each sample, expression of marker genes was normalized to
cyclophilin.
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).
Cells negative for all markers studied (expressing
cyclophilin only) were found primarily in the LOW and NEG populations (Figure 3c).
The data are summarized in Figure 2a — c. Values for the housekeeping gene
cyclophilin A were highly consistent across all cells analyzed.
Crystal structure of TM1367 from Thermotoga maritima at 1.90 A resolution reveals an atypical member of
the cyclophilin (peptidylprolyl isomerase) fold.
Expression of four stem cell marker genes normalized to
cyclophilin in nine single cell equivalents prepared from a pool of nine lysed HES3 cells.
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.
Data are normalized to
cyclophilin and expressed as ΔCt (marker gene —
cyclophilin).
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).
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).
That raised an intriguing question, because one of the few things scientists already knew about
cyclophilin A was that it is the target of a powerful immunosuppressant drug, cyclosporin A.
But
cyclophilin A is needed for so many other cellular activities that any treatment that inactivated it would probably be toxic.
The drug forms a complex with
cyclophilin A and a third protein, calcineurin, which is needed to activate the T cells of the immune system.
Jeremy Luban and his colleagues at Columbia University in New York, and Heinrich Göttlinger's team at Harvard Medical School in Boston, studied the interaction of HIV with
cyclophilin A in laboratory cultures.
Other primate immunodeficiency viruses, such as SIV and HIV - 2, the rarer and less virulent human virus, do not need their host's
cyclophilin A to replicate.
If
cyclophilin A could be somehow switched off, HIV could be controlled in the body.
A family of cis — trans peptidylprolyl isomerases that includes
cyclophilins and FK506 - binding proteins (FKBPs).
So far, our structures span protein families such as
cyclophilins, kinases and proteases as well as pathways for biosynthesis, salvage and metabolism of isoprenoids, fatty acids, phospholipids, pyrimidines, purines and polyamines.
Designed assays to evaluate novel drugs, with an emphasis on cellular pathways involving the enzymes poly - ADP - ribose polymerase, poly - ADP ribose glycohydrolase,
cyclophilins and sirtuins.