Figure 4.

The effects of DMB on HTLV-1 induced autophagosome accumulation. (A) MT2 cells were transfected with SC or H3. At 24 h after transfection, the cells were lysed for immunoblot assays. (B,C) Hela (B) or PMA-THP1 (C) cells were transfected with SC or H3. At 24 h after transfection, the cells were co-cultured with MT2 cells for another 24 h. Then the cells were washed with PBS three times to remove MT2 cells and lysed for immunoblot assays. (D) Hela cells were transfected with EGFP-LC3. At 24 h after transfection, the cells were transfected with SC or H3. At 24 h after transfection, the cells were co-cultured with MT2 cells for another 24 h. Then the cells were washed with PBS three times to remove MT2 cells and confocal microscopy analyses were performed. (E) The number of LC3 puncta per cell from images as in (D) was counted from n > 400 cells for each treatment over 3 repeats. The bars represent mean ± SD. ***p <0.001. (F) Hela cells were transfected with SC or H3. At 24 h after transfection, the cells were pretreated with control DMSO or 5 mM 3-MA for 6 h. Then the cells were incubated with MT2 cells for 24 h in the presence of control DMSO or 5 mM 3-MA as indicated. The cells were washed with PBS three times to remove MT2 cells and lysed for immunoblot assays. β-actin was used as a loading control in the immunoblot assays. The data were representative of three independent experiments.