Figure 3

Anti-proliferative effect of Sudemycin E on HTLV-1-transformed cell lines. (A) Cell viability was determined by MTT assay (Cell Titer 96® Non-Radioactive Cell Proliferation Assay, Promega) 72 h after Sudemycin (D1 or E) exposure. Data represent normalized mean ± SD; *p < 0.05 was considered statistically significant using a Mann–Whitney U-test. (B) Time-course analysis over 96 h using 1 μM of Sudemycin. Experiments were performed twice, in triplicate for each cell type, with DMSO used as control. Data represent the normalized mean ± SD. (C) Sudemycin E-induced expression of short spliceoforms encoding caspases 2 and 9 and MDM2. Exon-specific RT-PCR was carried out 72 h after Sudemycin E exposure. The oligonucleotides used have been previously described [36]. (D) Sudemycin E reduces Tax expression. Western blot analysis of Tax expression was carried out with a mouse monoclonal anti-Tax antibody (clone 474) 72 h after Sudemycin E exposure. Histograms represent the quantification of Tax signals normalized against beta-actin. (E) Sudemycin E decreases gene expression level of HBZ and the relative ratio sHBZ/unHBZ. Isoform-specific qRT-PCR analysis was carried out after 72 h of Sudemycin exposure. HBZ isoform expressions were normalized to hprt-1 transcripts used as internal control.