Figure 1.

Western blot probing for the expression of RyR by undifferentiated and differentiated C₂C₁₂ cells. C₂C₁₂ cells were lysed, and their proteins were denatured in sample buffer. Protein (60 μg) was loaded and separated by SDS-PAGE in a 4–12% polyacrylamide gradient gel. The primary antibody (MA3-916) directed against the RyR detected a high-molecular-mass band of ∼500–600 kDa with the use of peroxidase detection and a LumiGLO substrate kit (KPL) in differentiated cells, confirming the findings of Bennett and colleagues (1996). RyR signals could not be detected in undifferentiated cells because expression was below the detection level. Comparison of the RyR signals in undifferentiated and differentiated C₂C₁₂ cells clearly shows the strong increase in RyR after differentiation of C₂C₁₂ cells. Numbers indicate the positions and molecular masses of protein standards in kDa.