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. Author manuscript; available in PMC: 2018 Dec 1.
Published in final edited form as: Virology. 2017 Dec;512:95–103. doi: 10.1016/j.virol.2017.09.009

Figure 1. UL97 directs LIN52 phosphorylation at multiple sites.

Figure 1

(A) U-2 OS cells were transfected with expression plasmids for Flag-tagged LIN52 (Flag-LIN52) together with either an empty vector (EV), plasmids expressing HA-tagged UL97 (97), a UL97-L1 motif mutant (L1), a UL97 kinase-deficient mutant (KD), cyclin A2 (A2)/CDK2 (K2), cyclin E1 (E1)/CDK2, cyclin D2 (D2)/CDK4 (K4) or cyclin B1 (B1)/CDK1 (K1). Lysates harvested 48 hours after transfection were harvested and incubated with (+) or without (−) lambda protein phosphatase (l-PPase) prior to analysis by western blot with the indicated antibodies. The phosphorylation-dependent band shifts of LIN52 were detected in phosphate affinity (Phos-tag) gels. (B, C) Transfections were performed as in panel A except Flag-LIN52 alleles carrying single or multiple serine to alanine exchanges were also included.