FIG 1.

HBZ upregulates BDNF expression. (A) BDNF expression in Jurkat clonal cell lines stably expressing HBZ-S1 or carrying the empty vector. RT-PCR was performed using cDNAs prepared with gene-specific primers for BDNF, UBE2D2, and HBZ. (B) BDNF expression in uninfected and HTLV-1-infected T-cell lines. The upper panel shows levels of BDNF and UBE2D2 mRNAs analyzed by RT-PCR. RT-PCR was performed using cDNAs prepared with gene-specific primers for BDNF and UBE2D2. The lower panel shows the numbers of copies of HBZ mRNA per copy of UBE2D2 mRNA in the cell lines. cDNAs were prepared using gene-specific primers for HBZ and UBE2D2 for quantitative real-time PCR. The graph shows data obtained with RNAs identical to those used for the upper panel. (C) Western blot detection of BDNF in culture media of uninfected and HTLV-1-infected T-cell lines. The membrane in the upper panel was probed with a BDNF antibody. The lower panel shows the membrane stained with Ponceau S. (D) Western blot quantification of secreted mature and pro-BDNF from three independent experiments. Band densities were quantified using ImageQuant TL (GE Healthcare) and adjusted to the percentage of total mature and pro-BDNF (set to 100%). (E) Western blot detection of HBZ in HTLV-1-infected T-cell lines. The membrane in the upper panel was probed with an HBZ antibody. The lower panel shows the membrane stained with Ponceau S. No WCE, no whole-cell extract. (F) BDNF and HBZ expression in MT-2 cells transduced with negative-control (V1) and HBZ (V2) shRNA lentiviral vectors. The graph shows real-time PCR data from two independent transfection assays, with values normalized to that for V1 (set to 1).