FIG 5.

IBDV proteins accumulated around acidic compartments. DF-1 cells were transfected with pEGFP-N1-IBDV VP1, -IBDV VP3, -IBDV pVP2, and -IBDV PP (polyprotein) and pEGFP-N1 using Lipofectamine 2000 for 24 h. To stain the acidic compartments, cultures were incubated in medium containing 50 nM LysoTracker red (Molecular Probes) for 30 min; then nuclei were stained with Hoechst 33342 for 1 h. The distribution of proteins was observed by live cell imaging (green, viral proteins; red, acidic compartments; blue, nucleus). Serial pictures obtained from the video at the indicated time points are shown.