FIG 5.

Expression levels of HCMV MIE genes in PLS1KO cells were higher than those in the parental cell line. HEL, 36T-3, PLS1KO-A, and PLS1KO-B cells were infected with HCMV at an MOI of 0.1 PFU per cell. At 6 h (A) or 48 h (B) (for MIE) and 72 h (C) (for gB) p.i., the cells were fixed and then subjected to immunofluorescence staining with an anti-HCMV MIE antibody (A, B) and an anti-HCMV gB antibody (C), respectively. Nuclear and cytoplasmic fluorescence-positive cells were counted as MIE and gB antigen-positive cells, respectively, under a fluorescence microscope. Data represent means and standard deviations of three independent experiments. Data represent average relative values, and error bars indicate standard deviations. **, P < 0.01 by Student's t test. (D) HEL, 36T-3, and PLS1KO-A cells were infected with the HCMV AD169 strain at an MOI of 3 PFU per cell, and cell lysates were prepared at the indicated time points (hours). Equivalent amounts of lysates were subjected to SDS-PAGE. IB was performed using an anti-HCMV MIE antibody to detect endogenous IE1 and IE2. Detection of endogenous β-actin was performed as described in Fig. 1B and C. (E) Band intensity was quantified as described in Fig. 1B and C.