FIG 1.

The expression level and pattern of PLSCR1 induced by HCMV infection differs in HEL and OUMS-36T-3 (36T-3) cells. (A) Total cell lysates were prepared using RIPA buffer, and a total of 5 μg of total cell lysates were subjected to SDS-PAGE. Immunoblotting (IB) was performed using an anti-PLSCR1 antibody to detect endogenous PLSCR1 or an anti-actin antibody for endogenous β-actin. (B) HEL cells were infected with the HCMV AD169 strain at a multiplicity of infection (MOI) of 3 PFU per cell, and cell lysates were prepared at the indicated time points (hours postinfection). Equivalent amounts of lysates were subjected to SDS-PAGE. IB was performed using an anti-PLSCR1 antibody to detect endogenous PLSCR1 or an anti-MxA antibody to detect endogenous MxA. Then, endogenous β-actin, as a loading control, was detected by reprobing the same membrane with an anti-actin antibody (upper panel). Band intensity was quantified using NIH ImageJ software, and levels were normalized to those of the internal β-actin control (lower panel). (C) 36T-3 cells were infected with HCMV AD169, and cell lysates preparation, IB, and quantification of band intensity were performed as described in panel B.