Fig. 4

Analysis of the satellite cell population in teratomas obtained from Pax7+/+ and Pax7−/− ESCs. a, b Expression of mRNAs encoding satellite cell markers (Myf5, Cdh15) (for each genotype n = 6). Expression was related to the levels observed in 13.5 d.p.c. mouse embryo (E13.5; n = 3), and normalized to mRNA encoding β-actin (Actb). c, d Western blot analysis of Myf5 (for each genotype n = 3) and M-cadherin levels (for each genotype n = 3) in teratomas obtained from three Pax7+/+ ESC lines and three Pax7−/− ESC lines. Hsp90 level was used as a loading control. Graph represents optical density of Myf5 and M-cadherin bands compared to density of Hsp90 bands (optical density of Hsp90 was accounted as 100%, OdU optical density, arbitrary units). e Immunolocalization of Myf5+ satellite cells (green), laminin (red), and nuclei (blue) in teratoma muscles. Scale bar 10 μm. f Percentage of the myogenic cells (Myf5+ cells) in teratomas (for each genotype n = 16). g Calculation of the satellite cells (SC) expressing Myf5 in teratoma muscles (for each genotype n = 4). h Immunolocalization of laminin (red) and nuclei (blue) in teratoma muscles. Scale bar 100 μm. i Expression of Lama1 (coding laminin alpha 1) (for each genotype n = 9). Expression was related to the levels observed in 13.5 d.p.c. mouse embryo (E13.5, n = 3), accounted as 100%, and normalized to mRNA encoding β-actin (Actb). White bars—values for teratomas obtained from Pax7+/+ ESCs, gray bars—values for teratomas obtained from Pax7−/− ESCs. Data characterized by non-normal distribution (g) was normalized. Data are presented as mean ± SD. Stars represent results of Student’s unpaired two-tailed t test: *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001