Fig. 2. Induction of CCND1 by VRK1. (A) Cytoplasmic (Cyt.) and nuclear (Nuc.) lysates from HeLa cells transfected with control vector (vector, pDsRed1-C1) or pDsRed1-C1-VRK1 were immunoblotted with the indicated antibodies. (B) HeLa cells transfected with mock vector (pFlag-CMV2), control siRNA (siCont), siVRK1 or siVRK3 were fractionated into cytoplasmic and nuclear lysates and then immunoblotted with the indicated antibodies. (C) Total protein lysates from HeLa cells transfected with control siRNA (siCont) or siVRK1 for the indicated periods were immunoblotted with the indicated antibodies. (D) HeLa cells transfected with control vector (vector, pFlag) or pFlag-VRK1 were treated with DMSO (vehicle), MG132 or actinomycin D (ActD) for 5 hours, followed by immunoblotting (IB) with the indicated antibodies and northern blotting (NB) with an antisense CCND1 probe. Ethidium bromide (EtBr) stained 18S and 28S ribosomal RNA (rRNA) was used as a loading control.