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. Author manuscript; available in PMC: 2008 Nov 1.
Published in final edited form as: Microbes Infect. 2007 Sep 14;9(14-15):1581–1589. doi: 10.1016/j.micinf.2007.09.008

Figure 4. Promoter activity of HIV-1 3’LTR and STAT3 expression in the lymphoma.

Figure 4

(A) Promoter activity of HIV-1 3’LTR by reporter assay. Schematic representation of promoter constructs used in transient transfection assays is shown on the left. Forty-eight hours after transfection, cells were collected and the luciferase activity was measured. The percentage relative luminescence units (RLU) were calculated by dividing firefly activity by renilla activity. Horizontal bars indicate standard deviations of three independent experiments. (B and C) No methylation in a promoter enhancer region of HIV-1 3’LTR in the HIV-1-integrated lymphoma. (B) CpG sites in the promoter enhancer region of 3’LTR of the HIV-1 provirus in the patient with HIV-1-integrated lymphoma (218-529 in GenBank DQ117603). CpG sites are in boldface and numbered from the 5’ end of the LTR (1–11). Nuclear factor-κB and Sp1 sites identified with Motif Search (Kyoto University Bioinfomatics center, Kyoto, Japan, http://motif.genome.jp/) at a 75% cut-off value are indicated by boxes with broken and solid lines, respectively. Sequences used for primers are indicated by underlining. (C) Levels of CpG methylation of the promoter enhancer region of HIV-1 3’LTR in the HIV-1-integrated lymphoma and lymph nodes in the patient. Results of bisulfite genomic sequencing coupled with TA cloning are shown. The methylation status of 10 clones for each sample is presented; methylation of each CpG site is expressed as a filled circle, and unmethylated sites are shown as open circles. Top, schematic description of CpG sites in the 3’LTR of (B). (D-F) Immunohistochemistry of STAT3. The HIV-1-integrated lymphoma cells expressed STAT3 predominantly in the nucleus (D), however, signals of STAT3 were weak and localized in the cytoplasm in the other case of KSHV-positive, AIDS-related lymphoma (E), and were very weak in a case of EBV-positive, AIDS-related lymphoma (F). Original magnification is x400.