Figure 4. Pluripotency of sorted Sca-1^high+ cells.

(A) Quantitative relative expression of mRNAs coding for cardiomyocyte specific genes (Nkx2.5, ventricular Myosin Light Chain, Troponin T, beta and alpha Myosin Heavy Chain) in cells cultured 11 or 21 days in two different media. Results expressed as fold-increase above the levels in cells directly after sorting, n = 4–8 different experiments per group. (B) Representative pictures of cells cultured 21 days in medium B and expressing α actinin and Troponin I proteins. (C) Representative immunostainings of cells cultured 7 days in medium A and stained for the expression of CD31, von Willbrand Factor (vWF) and smooth muscle actin. (D) Quantitative relative expression of mRNAs coding for cardiomyocyte specific genes in cells cultured 21 days in differentiating medium C with or without BNP. Results expressed as fold-increase above the levels in untreated cells. n = at least 7 different experiments. (E) Representative immunostainings of cells cultured 21 days in medium C with BNP and stained with Troponin I (green) antibody and DAPI (blue). (F) The number of cells positive for Troponin I were related to the number of nuclei. n = 4–7 different experiments and in total, 1313 cells in BNP treated cells and 360 cells in untreated cells were evaluated for the Troponin I expression. (A,D,F) All the results are means ± SEM, *p < 0.05.