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Caption: Figure 1: Serpinel gene (PAI-1) expression is higher in irradiated arteries (XRT) vs unirradiated arteries (0XRT; ddCt), in relation to PGK1 (da), p = 0.012.
A decrease in [O.sub.2] level inhibits PHD, which in turn leads to HIF stabilization and transactivation of its target genes, for example, PGK1 or VEGF .
Concurrently, PGK1, another important enzyme at ATP generation step in glycolysis, has been reported by us to be downregulated by PPAR gamma upon activation by 15d-[PGJ.sub.2] .
MVIH upregulation was shown to be an independent risk factor in predicting poor RFS, and subsequent research indicated that MVIH may activate angiogenesis by inhibiting phosphoglycerate kinase 1 (PGK1) secretion.
As expected, proteins belonging to the same pathway (e.g., glycolysis) show overall good correlation (Figure S1.D, red CV% <35%), reflecting their regulated coexpression, but they also show quantitatively stable expression with other functional pathways: for example, the molar expression level of the Krebs cycle enzyme malate dehydrogenase 2 (MDH2) is relatively stable not only towards enzymes of the upstream glycolysis (ALDOA, TPI1, GAPDH, and PGK1), but also to the amount of downstream mitochondrial F0F1 ATPase (ATP5A1), and even to unrelated functional pathways such as proteins synthesis/turnover (ubiquitin C, UBC, and heat shock protein A8, HSPA8), cytoskeleton (profilin 1, PFN1), and cell signaling (calmodulin 1, CALM1).
The data is presented as relative expression to the mean of [beta]-D-glucoronidase (GUSB), peptidylprolyl-isomerase-A (PPIA), and phosphoglycerate-kinasel (PGK1) reference gene expression and to the expression at day 0.
Use of the RecoverAll kit produced RNA of higher quality (according to the [A.sub.260]/ [A.sub.280] ratio) and more reproducible RT-PCR amplification of a housekeeping gene (PGK1, phosphoglycerate kinase 1) than guanidinium thiocyanate-phenolchloroform extraction.
SAGE analysis revealed the expression changes of genes related to lipid metabolism [crystalline, zeta (Cryz); phosphatidic acid phosphatase type 2c (PPap2c)], carbohydrate metabolism [phosphoglycerate kinase 1 (Pgk1); sorbitol dehydrogenase 1 (Sdh1)], and amino acid metabolism (glutamate dehydrogenase (Glud); ornithine decarboxylase, structural (Odc).
 Human genes: RPLPO, ribosomal protein, large, P0; PPIA, peptidylprolyl isomerase A (cyclophilin A); TNFRSF1A, tumor necrosis factor receptor superfamily, member 1A; CCL11, CCL17, CCL19, CCL22, CCL3, CCL4, chemokine (C-C motif) ligand 11, 17, 19, 22, 3, and 4; CL5 chemokine (C-C motif) ligand 5; CCR1, CCR7, CCR8, chemokine (C-C motif) receptor 1, 7, and 8; CD86, CD86 molecule; CXCL12, chemokine (C-X-C motif) ligand 12 (stromal cell-derived factor 1); CXCL2, chemokine (C-X-C motif) ligand 2; CXCR4, chemokine (C-X-C motif) receptor 4; L128, interleukin 12B (natural killer cell-stimulating factor 2, cytotoxic lymphocyte maturation factor 2, p40); IL15, IL4, IL8, interleukin 15, 4, and 8; HPRT1, hypoxanthine phosphoribosyltransferase 1; PGK1, phosphoglycerate kinase 1.
Meisenhelder et al., "Mitochondria-translocated PGK1 functions as a protein kinase to coordinate glycolysis and the TCA cycle in tumorigenesis," Molecular Cell, vol.
We also tested 5 housekeeping genes [LMNB1, lamin B1; EIF1AX, eukaryotic translation initiation factor 1A, X-linked; CASC3 (also known as MLN51), cancer susceptibility candidate 3; PPIA; and PGK1, phosphoglycerate kinase 1] as endogenous controls (data not shown).
(b)-(d) The hypoxia environment under 1% [O.sub.2] culture was confirmed by the elevated expression of HIF1[alpha], PGK1, and Glut1; ** p < 0.01; n = 3.
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- PGI2-stimulating factor