Fig. 5
From: An adult myogenic cell line of the Japanese fire-bellied newt Cynops pyrrhogaster
Changes in immunoreactivity for Myf5, MRF4 and myogenin during the differentiation of CpM01 into myotubes. Blue: DAPI nuclear stain. (a–c) Representative immunofluorescent images of cells before induction (left column; n = 3) and at 6 days after induction (right column; n = 3). Scale bars, 200 μm. (d–f) Immunofluorescence intensity of the nucleus before induction and at 6 days after induction. In each condition, the mean luminance of fluorescence in the nucleus was individually measured for all cells in six randomly captured images of rectangular (726 μm × 544 μm) fields in three dishes after immunostaining (see Methods). Note that perinuclear and cytoplasmic fluorescence were not measured. For all three factors, the distribution patterns of the mean luminance of a nucleus changed significantly within 6 days after induction (two-sample Kolmogorov-Smirnov test; Myf5, n = 1894 (before), n = 2108 (after), P < 0.001; MRF4, n = 1419 (before), n = 1968 (after), P < 0.001; myogenin, n = 1888 (before), n = 2876 (after), P < 0.001). Note that for all three factors, nuclei exhibiting saturation levels of high immunofluorescence at 6 days after induction were localized in myotubes.
