Inefficient establishment of KSHV latency suggests an additional role for continued lytic replication in Kaposi sarcoma pathogenesis
J. Clin. Invest. Adam Grundhoff, et al. 113:124 doi:10.1172/JCI17803 [
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Figure 1Loss of TR-containing reporter plasmids from transfected cells. (
a) Functional elements of reporter plasmids. The vector backbone pGFP contains a GFP expression cassette driven by the CMV promoter. pGTR4 contains a GFP expression cassette as well as four contiguous units of the viral terminal repeats in authentic head-to-tail orientation. Construct pGTR4:73 contains, in addition to the GFP and TR elements, a CMV promoter–driven expression cassette for ORF73/LANA. (
b–
d) FACS analysis of cell lines transfected with the reporter plasmids described above. pGFP (open circles/dashed lines), pGTR4 (open squares/solid lines), or pGTR4:73 (filled triangles/solid lines) were introduced in SLK (
b), BCBL-1 (
c), or BJAB cells (
d and
e). BCBL-1 cells were only transfected with pGFP and pGTR4 because ORF73/LANA is provided
in trans by endogenous KSHV episomes. The percentage of GFP-expressing cells was monitored over a period of 13–15 days after transfection by flow cytometry (FACS). (
b–
d) Analyses of transfected mass cultures. For the data shown in
e, BJAB cells were FACS sorted 3 days after transfection for GFP expression in order to eliminate untransfected cells.