Gene transcription, ND10, and splicing compartments. (A) The plasmids used for making HSV-1 amplicons. (B) HEp-2 cells were transfected with pgfplacI; 6 hours later, the cells were super-infected with the amplicon ASK/E-Op for 2 hours. Then the cells were stained for ND10 using PML antibody and Texas Red-conjugated secondary antibody. Pictures were taken to show the relationship of ND10 and input amplicon DNA. (C) The same as (B), but the infection with the amplicon was at 12 hours, and the in situ hybridization experiment examined RNA (in blue), showing the relationship between lacZ gene transcription and ND10. (D) HEp-2 cells were infected with the amplicon (ASK/E-Op) for 2 hours; in situ hybridization was performed to show amplicon DNA (green). Immunofluorescence shows ND10 (red). (E) The same as (D), but the infection was for 12 hours; in situ hybridization was performed to show diffused β-gal RNA (DNA was degraded by DNAse treatment). (F) The same as (E), but the picture was taken to show the putative RNA. (G) HEp-2 cells were infected with the amplicon (ASK/E) and ICP0-deleted HSV-1 (as helper virus) for 12 hours; FISH was performed to show the distribution of lacZ RNA (green), ND10 (red), and splicing compartments (blue). (H) The same as (G) to show the relationship of RNA and splicing compartments. (I) The same as (G), to show the relationship of ND10 and RNA.