Fig 3. Involvement of SPARC in myoblasts fusion.

Myoblasts (4×10⁴ cells/well in 24-well plates) were induced to differentiate in DM with/without the induction/inhibition of SPARC for 5 days. The mediums were changed each 48 h. C2C12 were stained with May-Grünwald and Giemsa solutions, and the stained wells were observed under light microscopy. One-way ANOVA followed by the Tukey's HSD post-hoc test was used. All results are reported as means ± SEM. The addition of rSPARC (2 μg/ml) induced myotube formation, whereas, anti-SPARC (10 μg/ml) inhibited it. (a) Captured microscopic images of C2C12 fusion. The images were captured using 30× objectives under light microscopy (see S2 Fig for 100× objectives). Examples of multinucleated myotubes were shown by arrows. (b) Fusion index was calculated from 10 different photos per condition. Abbreviations: PBS: phosphate buffered saline, rSPARC: recombinant SPARC protein, and anti-SPARC: anti-SPARC antibody. *Significant differences between experimental conditions (n = 3): p < 0.05.