Validation of Housekeeping Genes as Reference for Reverse-Transcription-qPCR Analysis in Busulfan-Injured Microvascular Endothelial Cells

Figure 5

The impact of the selected different housekeeping genes on the target gene expression. Fold change values of three genes (ctnnb1, robo4, and notch1) were calculated based on actb, gapdh, ywhaz, and alas1. When using ywhaz and alas1 as reference genes, which were selected as the best two candidate housekeeping genes, the fold change values did not greatly increase over time. When using actb and gapdh as reference genes, which were commonly used, very higher values of fold change were obtained, with strong variation during cell injury.