To further explore how HCMV regulates Akt activity, we infected telomerase-immortalized human fibroblasts with HCMV strain TB40/E at a multiplicity of infection (MOI) of 2 50% tissue culture infective doses (TCID50)/cell and used phosphospecific antibodies to monitor phosphorylation of Akt at two sites crucial for its kinase activity, Thr308 and Ser473, and of PRAS40 (proline-rich Akt substrate of 40 kDa) at Thr246, a canonical indicator of Akt kinase activity (24) (Fig. 1A and B). This evidence concerns the gene AKT1S1 and infection.