Figure 3.

Hypoxic preconditioning reduces apoptosis of mesenchymal stem cells/multipotent stromal cells (MSCs) in vitro. After hypoxic preconditioning for 48 hours and culture for further 9 days, as described in Figure 3, MSCs were stained with EthD-III (dead cells) and DAPI (total nuclei). (A): Representative images of EthD-III and DAPI staining. (B): Quantification of A (n = 5). (C): Also after 9 days in culture, cells were stained with Annexin V and measured by flow cytometry (n = 5). After 3 days in culture under serum deprivation and hypoxia, total proteins were extracted and expression of apoptosis-related proteins detected using an antibody-based array. (D): Representative images. (E): Quantification of significant differences detected in the antibody array (n = 4). (F): These results were further confirmed by Western blot (n = 3). Statistical analysis was performed by paired Student’s t test; *, p < .05.