Figure 4
Effects of IFN-α and AZT on HTLV-1 expression and cell growth of HTLV-1 infected cells. ILT-Hod and ILT-#29 cells (106/ml) were cultured in the absence or presence of IFN-α (3000 IU/ml) and/or AZT (10 μM) as indicated, and HTLV-1 expression (A), cell growth (B), cell cycle (C), and Ki-67 expression (D) in the cells were evaluated. A. Expression of intracellular Tax protein 3 days after the initiation of culture was evaluated by flow cytometry following stained with anti-Tax (open histogram) and isotype control (closed histogram) antibodies. B. ILT-Hod and ILT-#29 cells were similarly treated with IFN-α and/or AZT, and maintained with addition of equal volumes of fresh medium without IFN-α or AZT on the day 1 and 3, then viable (closed bar) and non-viable (open bar) cell numbers in cultures were evaluated by trypan blue exclusion on the day 8. *p < 0.05. C. ILT-Hod and ILT-#29 cells similarly treated with IFN-α and/or AZT were subjected to cell cycle analysis on the day 8. Cultures were treated with BrdU (10 μM) for the last 24 h of culture then permeabilized and incubated with a FITC-labeled mouse anti-BrdU antibody and 7AAD. Cells that are 7AAD-negative can be considered apoptotic (Apo). BrdU-negative and 7AAD-intermediate positive cells are in the G0/G1 phase. BrdU-positive and 7AAD-positive cells are in the S phase. BrdU-negative and 7AAD-highly positive cells are in the G2/M phase. The values in the dot plots indicate the proportion of the cells (%) in each phase. D. ILT-Hod and ILT-#29 cells similarly treated with IFN-α and/or AZT were analyzed for intracellular Ki-67 expression by flow cytometry on the day 8. The values in the dot plots indicate the proportion of Ki-67-positive cells (%).