Figure 4.
Cardiomyocyte differentiation of normoxic and hypoxic P2 CDCs. (A) The relative mRNA expression of Nkx 2.5, Tnnt, and MyHC for normoxic and hypoxic P2 CDCs after treatment with DMSO for 2 weeks under (i) normoxia (left panel) or (ii) hypoxia (right panel) conditions (n = 3). mRNA expression of 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. #p < 0.05 between the two groups linked with arrow. (B, C) Representative confocal images (of three independent experiments) of the DMSO differentiation protocol in normoxia (B) and hypoxia (C). 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, troponin T, or myosin heavy chain. Left panel: normoxic CDCs (control or treated with DMSO), right panel: hypoxic CDCs (control or treated with DMSO). Scale bars: 50 µm.