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. 1998 Sep 1;95(18):11020–11025. doi: 10.1073/pnas.95.18.11020

Figure 7.

Figure 7

CK2 phosphorylation is required for the formation of a DNA–protein complex containing CCA1. (A) Phosphatase treatment abolishes CCA1 binding. WCE were incubated in the absence (lane 1) or presence (lane 2) of λ protein phosphatase (200 units added to 20 μl) for 30 min at 30°C. (B) The CCA1-containing complex is more abundant in plants overexpressing CCA1 than in wild type. WCE from wild type (WT) and a CCA1-ox line (CCA1-ox) (7) grown in 12:12 photoperiods were used in the EMSA. (C) Inhibition of CK2 activity in plant extracts abolishes CCA1 binding. WCE were incubated with the indicated inhibitor for 45 min at 30°C. Lanes: 1, no inhibitors; 2, 5 mM 2,3-diphosphoglycerate (D.G.); and 3, 100 μM quercetin. Arrows indicate the position of the major DNA–protein complex containing CCA1.