Changes in ROS-related gene expression and ROS generation in hBM-MSCs over time. (a) An ROS-related gene network was constructed algorithmically by IPA. Red and green areas indicate up- and downregulated genes, respectively. Differentially expressed genes were identified by microarray analysis without fold-change cut-off. (b) Semi-quantitative RT-PCR detection of ROS-related genes (AKT2, MAP3K2, PIK3R1, FOXO3, and KHDRBS1) in hBM-MSCs after 6 or 12 h of storage in PBS. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as an internal control. Bands were cropped from Supplementary Fig. 2. (c) ROS-related gene expression in hBM-MSCs stored in PBS for 6 or 12 h, as determined by qPCR. Relative gene expression levels were normalized to that of GAPDH and compared to the level at 0 h (control). (d) Evaluation of intracellular ROS generation after 6 or 12 h using DCFH-DA. The intensity of non-oxidized DCFH-DA was used as a blank. Data represent mean ± SD of three independent experiments. *P < 0.05, **P < 0.001 vs. 0 h (control).