Fig. 4

EBV lytic cycle induction by TGF-β in latency I infected MUTU-1 cells. MUTU-1 cells were either uninduced (time zero) or treated with TGF-β (5 ng/mL) for the indicated time. a Immunoblots for IFI16 (top), ZEBRA (BZLF1; middle), or GAPDH (bottom) following induction. All bands were normalized to GAPDH with the fold change calculated relative to the uninduced sample. b EBV-negative BJAB cells were treated with TGF-β similar to MUTU-1 cells or left untreated and immunoblotted for IFI16, ZEBRA, and GAPDH. c Real time DNA PCR for the relative EBV genome abundance. Primers specific to the latent EBNA1 gene were used and the level of DNA was normalized against the β-tubulin level. d Real time qRT-PCR for mRNA levels in uninduced to 96 h post TGF-β induction in MUTU-1 cells. Total RNA was extracted and mRNA levels analyzed by real-time qRT-PCR using gene-specific primers to EBV temporal class genes. mRNA values were normalized against β-tubulin mRNA levels. All values are represented as the relative amount compared to the uninduced (time zero) control. Results are presented as ± SD of data from three independent experiments and statistical analysis was done with a Student’s t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant