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. 2010 Nov 5;9(21):4379–4386. doi: 10.4161/cc.9.21.13671

Figure 2.

Figure 2

FGF signaling strongly downregulates activity of cyclin B1/CDK1 complexes in chondrocytes. RCS cells were treated with FGF1 for indicated times. (A and C) Kinase activity of immunoprecipitated cyclin B1/CDK1 complexes was assayed in vitro. The cyclin B1/CDK1 complexes were isolated from 1 mg of total cellular protein using anti-cyclin B1 antibodies, and histone H1 was used as a substrate. Antimouse IgG was used as a negative control. An equal amount of protein loading was confirmed by immunodetection of cyclin B1 and CDK1 in immunoprecipitated complexes. (B) 20 µg of total cellular protein were analyzed by SDS-PAGE followed by immunoblotting for anti-phospho-CDK1(Y15), anti-CDK1 and anti-phospho-CDK1(T14) antibodies. Equal amount of protein loading was confirmed by α-tubulin and α-actin immunodetection. (C) RCS cells were pretreated with Nocodazole for 12 hrs and then FGF1 was added for indicated periods of time.