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. 2012 Sep 6;8(9):e1002908. doi: 10.1371/journal.ppat.1002908

Figure 1. Establishment of 4sU-tagging for lytic MCMV infection.

Figure 1

(A) Incorporation of 4sU throughout MCMV infection. Cells were infected with MCMV at an MOI of 10 and exposed to 200 µM 4sU for 1 h at different times of infection before total RNA was isolated. Thiol-specifically biotinylated RNA was subjected to dot blot analysis in 10-fold dilutions (1 µg down to 1 ng). A biotinylated oligonucleotide of 81 nt (PC, 100 ng down to 0.1 ng) was used to quantify 4sU-incorporation; M = mock control. (B)–(D) Comparison of genes identified to be regulated in newly transcribed RNA to genes regulated in total RNA. (B) Numbers of genes up- and down-regulated (>2-fold, p≤0.05) at different times of infection are shown for newly transcribed RNA and total RNA. (C) Venn diagrams of all genes regulated more then 2-fold in newly transcribed RNA and total RNA. (D) Venn diagrams showing genes regulated >2-fold in total RNA at 2, 4 and 6 hpi and in newly transcribed RNA at and prior to the indicated time point of infection; red = newly transcribed RNA, blue = total RNA.