Figure 7.
Effects of PHDI preconditioning on CDC cardiomyogenic differentiation. (A) The relative mRNA expression of Nkx 2.5, Tnnt, and MyHC for normoxic and PHDI-treated hypoxic P2 CDCs after treatment with DMSO for 2 weeks under normoxia (n = 3). mRNA expression of PHDI-treated hypoxic cells was normalized to the geometric mean of GAPDH and Actb (housekeeping genes) and nontreated control cells (calibrator). *p < 0.05 versus nontreated control. (B) Representative confocal images (of three independent experiments) of CDCs treated with DMSO or BIC for 24 h, followed by differentiation with DMSO for 2 weeks. An equal number of cells (8 × 105 per chamber) of each condition were trypsinized and seeded on four-well chamber slides coated with fibronectin (10 µg/ml) and stained with DAPI and α-sarcomeric actin or troponin T. Scale bars: 20 µm. All experiments in this study used P2 CDCs at 70% to 80% confluency, unless otherwise stated. Scale bars: 20 µm.