PKC-Mediated ZYG1 Phosphorylation Induces Fusion of Myoblasts as well as of Dictyostelium Cells

Figure 8

Enhanced formation of multinucleate cells by transfecting with the mzyg1 gene. The percentages of cells containing more than two nuclei are shown as fusion indexes. The pIRES2-AcGFP vector or pIRES2-AcGFP vector containing mzyg1 gene was transfected into C2C12 cells as described in Section 2. After 1 day, cells were fixed with 4% paraformaldehyde and mounted in PBS containing 20% glycerol and DAPI (final concentration: 2.5 μg/mL) after three washings with PBS. The numbers of cells and nuclei stained with DAPI were counted. Three samples in two independent experiments were counted (about 500~1,000 cells/sample). The fusion index (ratio of the cell number participated in cell fusion to the total cell number) was calculated by the following formula: Fusion index (%) = 2 × (The number of cells containing 2 nuclei) + 3 × (The number of cells containing 3 nuclei)⋯𝑛 × (the number of cells containing 𝑛 nuclei)/1 × (The number of cells containing 1 nucleus) + 2 × (The number of cells containing 2 nuclei) + 3 × (the number of cells containing 3 nuclei⋯𝑛 × (The number of cells containing 𝑛 nuclei) × 100. GFP; C2C12 cells transfected with the pIRES2-AcGFP vector. GFP and ZYG1; C2C12 cells transfected with the pIRES2-AcGFP vector containing the mzyg1 gene. Bars: standard deviations.