Fig. 2.

RPL4 is essential for EBNA1 transcription activation and binding of oriP and LCL growth. (A) BJAB cells were transduced with lentiviruses expressing short hairpins targeting RPL4, shRPL4 #1 or shRPL4 #2, or Scr.. RPL4 depletion was shown by Western blot. These cells were then transfected with EBNA1 expression plasmid and oriP luciferase reporter plasmid and used to assay for EBNA1-dependent transcription of the oriP-Luc reporter plasmid. Luciferase activities were represented as mean ± SD from three independent experiments here and all of the following transfection luciferase reporter assays described elsewhere in this article. (B) RPL4 was depleted from LCL1 with shRNA. Antibodies against EBNA1 or control were used in ChIP assays to precipitate oriP DNA from LCLs. oriP DNA precipitation was quantitated by qPCR and presented as % of input DNA (mean ± SD) from three independent experiments and for all following ChIP experiments. (C) EBV copy number was determined by qPCR in LCL depleted for RPL4 or shRNA control, with untreated LCL set to 1. DNA was normalized to GAPDH genomic DNA. (D) Cells were counted for 5 d after shRNA knockdown. Dead cells were excluded by trypan blue staining.