Figure 2.

Galectin-3 promotes myogenesis. A) Myogenesis was induced in the presence or absence of different concentrations of lactose, an antagonist of galectin-3, for 72 h. Cells were fixed and stained with anti-MHC antibody and DAPI. The total areas of MHC⁺ multinucleated myotubes were calculated using in-house software, and the fold increase in myogenesis was calculated with cells treated with PBS(−) as 1. Each data point represents the mean of the fold increase obtained by 3 independent experiments with error bars indicating sd. Significance compared with control (0 mM) by 1-way ANOVA is shown. *P < 0.05, **P < 0.01. When ANOVA–on ranks was used, the detected significance was as follows: lactose 5 mM. B, C) Myogenesis was induced in the presence or absence of galectin-3 for 62 h (B) and 72 h (C). The number of nuclei in MHC⁺ and multinuclear myotubes were counted and the fold increase in myogenesis was calculated taking cells treated with PBS(−) as 1. Each data point represents the mean of the fold increase obtained from 3 independent experiments with error bars indicating sd. Significance compared to control (0 µM) by 1 way -ANOVA is shown. *P < 0.05, **P < 0.01. When ANOVA–on ranks was used, the detected significance was the following: galectin-3, 0.2 µM and 2 µM. C) The number of nuclei in each MHC⁺ multinucleated myotube found in the stitched FOVs was counted and categorized into 3 groups based on the number of nuclei in each myotube. The representative results obtained from 2 independent experiments are shown. D, E) Stitched images (7 × 5 FOVs) of extended-focus images of myotubes (MHC⁺ cells are in white) formed in the absence (D) and presence (E) of galectin-3 after differentiation for 72 h are shown together with the size of 1 FOV. Representative images of 3 independent experiments are shown.