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. 2008 May 1;105(18):6584–6589. doi: 10.1073/pnas.0802785105

Fig. 1.

Fig. 1.

The cellular levels of Mad1 are rapidly reduced upon mitogen or insulin stimulation. (A) HeLa cells were cultured in the presence (S) or absence of serum (SD) for 48 h before stimulation with serum (SS) for the indicated amount of time. The lysates were fractionated and analyzed by Western blotting with the indicated antibodies. (B) HeLa cells were starved for 48 h and pretreated with DMSO or MG132 (10 μM) for 1 h before serum stimulation for 3 h. The Mad1 levels were analyzed by Western blotting. (C) The experiment was performed as in A except by using HeLa-Flag-Mad1 cells and stimulating with insulin (10 μg/ml) as indicated. (D) HeLa-Flag-Mad1 cells were starved overnight and pretreated with DMSO or MG132 (10 μM) for 1 h before stimulation with insulin (10 μg/ml) for 30 min. The Mad1 levels were analyzed by Western blotting. (E) HeLa cells were starved (SD) with serum-free medium for indicated amount of time. The cell lysates were analyzed by using Western blotting with the indicated antibodies.