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. 2013 Jul 29;288(37):26678–26687. doi: 10.1074/jbc.M113.466433

FIGURE 2.

FIGURE 2.

Cyclin D1 interacts with NDR1/2 independent of Cdk4. A, 293T cells were cotransfected with pCMV-Myc-cyclin D1 alone or with pCMV-FLAG-NDR1 or pCMV-FLAG-NDR2. The cell lysates were immunoprecipitated (IP) with anti-FLAG antibody and then immunoblotted (IB) with anti-Myc antibody. B, 293T cells were cotransfected with pcDNA3-Cdk4 alone or with pCMV-Myc-cyclin D1 or pCMV-Myc-cyclin D1 K112E. The cell lysates were immunoprecipitated with anti-Myc antibody and then immunoblotted with anti-Cdk4 antibody. C, 293T cells were cotransfected with pCMV-FLAG-NDR2 alone or with pCMV-Myc-cyclin D1 or pCMV-Myc-cyclin D1 K112E. The cell lysates were immunoprecipitated with anti-Myc antibody and then immunoblotted with anti-FLAG antibody. D, GST protein or GST-NDR2 immobilized on glutathione beads was incubated with His-cyclin D1 in lysis buffer. The associated protein was eluted with SDS loading buffer and immunoblotted with anti-cyclin D1 antibody. TCE, total cell extract; CBB, Coomassie Brilliant Blue.