Journal: The Plant Journal

Article Title: The Arabidopsis cyclin-dependent kinase-activating kinase CDKF;1 is a major regulator of cell proliferation and cell expansion but is dispensable for CDKA activation

doi: 10.1111/j.1365-313x.2009.03884.x

Figure Lengend Snippet: Figure 5. The activities of cyclin-dependent kinase activating kinases CDKA and CDKB are not reduced in the cdkf;1-1 mutants. (a), (b) Protein levels and kinase activities of CDKA and CDKB in the wild-type and the cdkf;1-1 mutants. Protein extracts (10 lg for CDKA, 40 lg for CDKB and 5 lg for actin) from 10-day-old seedlings (a) or calli (b) were subjected to immunoblotting with specific antibodies. A kinase assay was conducted with immunoprecipitates by using histone H1 as a substrate. (c) Detection of the phosphorylated form of CDKA;1. Protein extracts (60 lg) from calli were incubated with or without lambda protein phosphatase (left), or depleted with anti-CDKA;1 antibody (right). Then the samples were subjected to immunoblotting with anti-CDKA;1 or anti-phospho-Cdc2 (Thr161) antibody. CDKA-P indicates CDKA;1 phosphorylated within the T-loop. (d) Protein extracts (60 lg) from calli of wild-type plants and the cdkf;1-1 mutants were subjected to immunoblotting with anti-phospho-Cdc2 (Thr161) antibody. Bands with an asterisk representing non-specific cross-reactions with the antibody.

Article Snippet: Commercial antibodies used include anti-actin antibody (MP Biomedicals, http:// www.mpbio.com/) and anti-phospho-Cdc2 (Thr161) antibody (Cell Signaling Technology, http://www.cellsignal.com/).

Techniques: Western Blot, Kinase Assay, Incubation