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. 2010 Aug 15;21(16):2966–2974. doi: 10.1091/mbc.E10-01-0015

Figure 1.

Figure 1.

HIPK2 interacts with CREB and induces CREB retardation on SDS-PAGE. (A) Left, 293 cells were transfected with HA-CREB alone, HA-CREB plus Flag-HIPK2 wild type (WT), or HA-CREB plus Flag-HIPK2 kinase dead (KD). Whole cell lysates prepared after 48-h incubation were immunoprecipitated with anti-HIPK2 antibody, and the immunoprecipitates were subjected to Western blotting with anti-HA antibody (top). The whole cell lysates were also analyzed by Western blotting for expression of transfected HA-CREB (middle) and HIPK2 (bottom). Right, whole cell lysate of nontransfected 293 cells was immunoprecipitated with goat IgG or anti-HIPK2 antibody, followed by Western blotting with anti-CREB antibody. pCMVhumanCREB-transfected cell lysate was loaded as a CREB positive control. (B) K562 cells were transfected with nontagged CREB, or CREB plus Flag-HIPK2-WT, and whole cell lysates were treated with protein phosphatase (PPase +) before SDS-PAGE and Western blotting with anti-CREB antibody.