FIG 6.

PLSCR1 specifically interacts with HCMV IE1 and IE2. HEK-293 cells (1 × 10⁶) were plated in 60-mm cell culture plates and transfected with 2 μg of pcDNA3 or S-PLSCR1 and 0.5 μg of pUCIE1 or 3 μg of pUCIE2. A total of 300 μg of total cell lysate prepared in NP-40 lysis buffer was incubated with S-protein beads to precipitate PLSCR1. Following pulldown assays, 1 μg (IE1) or 5 μg (IE2) of total cell lysate and the precipitated complexes were divided into two parts and subjected to SDS-PAGE. IB was performed using an anti-S tag antibody to detect PLSCR1 or an anti-HCMV MIE antibody to detect IE1 and IE2.