Figure 2.

eEF2 contains symmetrically dimethylated arginines induced by bFGF. (A) bFGF induced the level of symmetric arginine dimethylation but neither asymmetric arginine dimethylation nor expression of eEF2. NIH3T3 cell lysates were immunoprecipitated with anti-eEF2 antibody, and immunoprecipitation with normal goat anti-rabbit IgG was performed as a negative control. Western blotting was performed with anti-eEF2 antibody, SYM10 and ASYM24, and protein bands were visualized using enhanced chemiluminescence. GAPDH was used as a loading control. In put, whole cell lysates; IP, immunoprecipitation; WB, western blot. (B) The symmetrically dimethylated arginine-containing 95-kDa protein is eEF2. bFGF induced symmetric arginine dimethylation of eEF2 but did not affect the level of eEF2 expression. NIH3T3 cell lysates were prepared, immunoprecipitated with SYM10, and analyzed using anti-eEF2 antibody and SYM10 Western blots, respectively as in (A).