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Figure 6


Fig. 6. VRK1 phosphorylates CREB at Ser133. (A) Phosphorylation of CREB by wild-type or kinase-dead VRK1 was assessed by in vitro kinase assays. GST-CREB-fragments (F1-F4), wild-type CREB or site-directed mutated CREB (S133A and S142A) proteins were used as substrates. (B,C) Cytoplasmic and nuclear lysates from HeLa cells transfected with kinase-dead (KD) or wild-type (WT) VRK1 (B), or with control siRNA (siCont) or siVRK1 (C) were immunoblotted with the indicated antibodies. Protein disulfide isomerase (PDI) and Lamin B were used as fraction markers for the cytoplasm and nucleus, respectively. (D,E) HeLa cells transfected with WT-VRK1 or KD-VRK1 were immunostained for CREB-P S133 (green) (D) or CREB (E). Scale bars: 20 µm.





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