Selection of Reference Genes for Normalization of Gene Expression After Exposure of Human Endothelial and Epithelial Cells to Hypoxia.

ABSTRACT: The selection of a stably expressed reference gene is a critical step for the quantitation of gene expression by qRT-PCR. We tested the stability of expression of nine putative reference genes in normoxia and hypoxia in four different human cell types: coronary (HCAECs) and pulmonary endothelial cells (HPAECs), EA.hy926 endothelial cells, and A549 alveolar epithelial cells. Cells were cultured in normoxic and hypoxic conditions for up to 72 h. Total RNA was isolated and used for qRT-PCR. Stability of expression was assessed by calculating the coefficient of variation of the cycle threshold (Ct CV) by pairwise comparison of ΔCt values, and by the NormFinder algorithm. A final rank was calculated for each gene. Finally, we analyzed VEGFA expression by using GAPDH or the optimal

SUBMITTER: Hannemann J 

PROVIDER: S-EPMC11855034 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature