FIG. 3.

Cdk9 recruitment to the viral transcriptosomes is delayed during infection at a high MOI with the IE2 86 ΔSX-EGFP virus. G₀-synchronized cells were released into G₁, infected with HCMV IE2 86 ΔSX-EGFP or IE2 86 wt-EGFP virus at an MOI of 5, or treated with mock supernatant and were seeded onto glass coverslips. At 8 (panels 1 to 6) and 16 (panels 7 to 15) h p.i., cells were washed with PBS, fixed with 2% FA, permeabilized, and stained with anti-cdk9 and Cy3-conjugated secondary antibodies. Nuclei were stained with Hoechst dye. The brightness of the image elements representing the intensity of EGFP expression in panels 4 to 6 was increased by use of the Softworx program for visualization purposes. Mock (M) infected samples at 16 h p.i. are represented by panels 13 to 15. The white arrows point to examples of colocalization. All of the images represent confocal optical 0.2-μm sections at a magnification of ×1,000 under conditions of oil immersion. wt, IE2 86 wt-EGFP virus; ΔSX, IE2 86 ΔSX-EGFP virus.