Versican V3 overexpression decreases p38 MAPK activation and does not alter JNK or ERK5 activation. A, subconfluent cells were treated with 25 ng/ml EGF in serum-free medium for the indicated times. After the treatment, cell extracts were prepared as described under “Experimental Procedures” followed by immunoblotting with anti-phospho-p38MAPK, anti-phospho-JNK1/2/3, and anti-phospho-ERK5 antibodies or anti-non-phosphorylated forms antibodies (as a loading control). B, densitometric analyses of the ratio of the area of the phosphorylated versus total kinases from LXSN and LV3SN MeWo cells. In the case of JNK, both bands corresponding to JNK1 and JNK2/3 were considered. A representative experiment out of four performed is shown. Values are mean ± S.D. *, p < 0.05 (versus t = 0 min).