Figure 1.

Interaction and, phosphorylation of Nek7 with RGS2. (A) Western Blotting (WB) analysis from pull-down (PD) of recombinant Nek7 binding to endogenous HEK293T RGS2. RGS2 does not bind to Ni-NTA agarose resins (Ni-NTA), nor does RARA bind to Nek7, demonstrating the assay specificity. The results are based on 3 independent experiments. Molecular weight (kDa) of the proteins is indicated. (B) Nek7 phosphorylates RGS2 in vitro. The arrowheads indicate the positions of the GST-tagged proteins or GST-control whereas the arrows indicate the position of the 6xHis-Nek7FL detected in the autoradiography (³²P Autorad), WB or in SDS-PAGE. The GST-NEK9(764–976) was used as a phosphorylation positive control.²⁴ No activity of 6xHis-Nek7FL was detected on GST-control, demonstrating the phosphorylation specificity for substrate. The results are based in 3 independent experiments. Molecular weights (kDa) of the marker proteins are indicated.