Figure 5.

Analysis of myotubes formation in C2C12 cells. Cells were seeded on gold homogeneous surfaces (a) or nanopatterned surfaces (b) and cultured to allow myotube formation. Immunofluorescence microscopy images show myosin heavy chain (MHC) staining of multinucleated myotubes (green) and DAPI nuclei staining (blue). Myotube formation is observed when cells are cultured on control surfaces in absence of BMP-6 (No BMP-6). For iBMP-6 and sBMP-6, cells were cultured in presence of 80 ng (A) or 19, 6 and 1 ng (B) of the growth factor. MHC positive areas were quantified by image analysis. In the graphs error bars are SD, # p < 0.05, § p < 0.01, * p < 0.001. Student’s t-test was used for single comparison.