Figure 5. Knockdown of Mi-2 accelerates the expression of muscle-specific proteins.

(5a) On day -1, C₂C₁₂ cells proliferating in growth media were transfected with Mi-2 or non-targeting siRNA. The next day (day 0), myoblasts were harvested or placed in differentiation media. At days 1, 2, 3, 4, and 5 protein lysates were prepared from differentiating cells. Equal amounts of protein were loaded and immunoblotted with antibodies against Mi-2, MyHC, myogenin, and vinculin. Mi-2 levels are reduced on the first day after transfection with Mi-2 siRNA (day 0). Myogenin and MyHC expression are accelerated by one day in the cultures treated with Mi-2 siRNA. This experiment was performed 4 times with similar results. (5b) Human myoblasts were placed in differentiation media at day 0 and harvested at days 1, 2, 3, 4, 5, 6, and 8. Equal amounts of protein were loaded and immunoblotted with antibodies against Mi-2, MyHC, and vinculin.