Cells were washed with PBS thrice, stained with 1% nuclear fast red to stain the cell nuclei, washed, air-dried, dehydrated, cleaned, and mounted for light microscope observation.
After washing steps and Hoechst staining of nuclei, the specimens were mounted for a confocal microscope examination.
Immunohistochemical studies were performed using the UltraSensitive S-P Kit (Maixin Biotechnology Company, Fuzhou, China) and color was developed using DAB as chromogen, counterstained with Mayer's hematoxylin and mounted for evaluation using microscope (OLYMPUS BX-51, Osaka, Japan).
Finally, slides were counter-stained with hematoxylin, fixed with ethanol, and cover-slipped with Eukitt mounting medium for microscope preparations (O. Kindler GmbH, Friburg, Germany).
Sections were then washed and mounted for observation under a scanning confocal microscope (TCS SP2 Leica).
The samples were rinsed with PBS and mounted for imaging with a Nikon TE2000 microscope.
They were finally counterstained with DAPI and mounted for examination using a Nikon confocal microscope (Eclipse Ti, Nikon, Japan).
Coverslips were mounted for viewing on an Olympus FV1000 confocal microscope.
Samples were then washed with blocking solution and mounted onto microscope slides for imaging.
Myoblasts, upon reaching 80% confluence, were trypsinized and mounted on microscope slides for homogeneity estimation with IHC staining using the myoblast-specific myosin heavy chain-1 (MHC-1) antibody (Sigma, M8421) Briefly, following trypsinization the cells were suspended in 1% bovine serum albumin (BSA)/PBS.
