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Reference genes for expression studies in hypoxia and hyperglycemia models in human umbilical vein endothelial cells.

Bakhashab S, Lary S, Ahmed F, Schulten HJ, Bashir A, Ahmed FW, Al-Malki AL, Jamal HS, Gari MA, Weaver JU - G3 (Bethesda) (2014)

Bottom Line: All five genes displayed stable expression under hyperglycemia.However, only RPLP0 and TFRC genes were stable under hypoxia up to 12 hr.Under hyperglycemia combined with hypoxia up to 12 hr, the expression of RPLP0, TFRC, GUSB, and ACTB genes remained unchanged.

View Article: PubMed Central - PubMed

Affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle NE2 4HH, United Kingdom Biochemistry Department, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia Center of Excellence in Genomic Medicine Research, King Abdulaziz University, P.O. Box 80216, Jeddah 21589, Saudi Arabia.

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Effect of hyperglycemia and hypoxia on gene expression of the selected reference genes. Fold change in gene expression analyzed by the 2−∆CT method. Data are mean ± SEM, n = 3–4, *P < 0.05 vs. control. Expression of GUSB, TFRC, RPLP0, and ACTB was found to be stable, whereas GAPDH was increased with borderline significance after 12 hr of hypoxia compared with the control.
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fig3: Effect of hyperglycemia and hypoxia on gene expression of the selected reference genes. Fold change in gene expression analyzed by the 2−∆CT method. Data are mean ± SEM, n = 3–4, *P < 0.05 vs. control. Expression of GUSB, TFRC, RPLP0, and ACTB was found to be stable, whereas GAPDH was increased with borderline significance after 12 hr of hypoxia compared with the control.

Mentions: When HUVEC were treated under hyperglycemic conditions (16.5 mM glucose) for 24 hr, the five selected reference genes demonstrated stable expression (Figure 3). Under hyperglycemia combined with hypoxia, gene expression of GUSB, TFRC, RPLP0, and ACTB turned out to be stable (Figure 3) in all time intervals measured (1, 3, and 12 hr). In contrast, an increase in gene expression of GAPDH was revealed with a borderline significance (P = 0.05) after 12 hr of hypoxic incubation compared with the control.


Reference genes for expression studies in hypoxia and hyperglycemia models in human umbilical vein endothelial cells.

Bakhashab S, Lary S, Ahmed F, Schulten HJ, Bashir A, Ahmed FW, Al-Malki AL, Jamal HS, Gari MA, Weaver JU - G3 (Bethesda) (2014)

Effect of hyperglycemia and hypoxia on gene expression of the selected reference genes. Fold change in gene expression analyzed by the 2−∆CT method. Data are mean ± SEM, n = 3–4, *P < 0.05 vs. control. Expression of GUSB, TFRC, RPLP0, and ACTB was found to be stable, whereas GAPDH was increased with borderline significance after 12 hr of hypoxia compared with the control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4232541&req=5

fig3: Effect of hyperglycemia and hypoxia on gene expression of the selected reference genes. Fold change in gene expression analyzed by the 2−∆CT method. Data are mean ± SEM, n = 3–4, *P < 0.05 vs. control. Expression of GUSB, TFRC, RPLP0, and ACTB was found to be stable, whereas GAPDH was increased with borderline significance after 12 hr of hypoxia compared with the control.
Mentions: When HUVEC were treated under hyperglycemic conditions (16.5 mM glucose) for 24 hr, the five selected reference genes demonstrated stable expression (Figure 3). Under hyperglycemia combined with hypoxia, gene expression of GUSB, TFRC, RPLP0, and ACTB turned out to be stable (Figure 3) in all time intervals measured (1, 3, and 12 hr). In contrast, an increase in gene expression of GAPDH was revealed with a borderline significance (P = 0.05) after 12 hr of hypoxic incubation compared with the control.

Bottom Line: All five genes displayed stable expression under hyperglycemia.However, only RPLP0 and TFRC genes were stable under hypoxia up to 12 hr.Under hyperglycemia combined with hypoxia up to 12 hr, the expression of RPLP0, TFRC, GUSB, and ACTB genes remained unchanged.

View Article: PubMed Central - PubMed

Affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle NE2 4HH, United Kingdom Biochemistry Department, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia Center of Excellence in Genomic Medicine Research, King Abdulaziz University, P.O. Box 80216, Jeddah 21589, Saudi Arabia.

Show MeSH
Related in: MedlinePlus