LMP1 mediates RON expression. EBV-negative Akata cells were electroporated with plasmids expressing EBNA1, LMP1, LMP2A, Zta, or Rta, as indicated. Total RNA and protein were harvested from each transfectant at 72 hours posttransfection. (A) RON transcripts were measured by RT-Q-PCR and the relative fold increase was normalized to the amounts of RON transcripts in vector control cells. (B) Expression of RON, EBNA1, LMP2A, Rta, LMP1, Zta, and GAPDH was detected by Western blotting. (C) EBV-negative Akata cells were infected with pSIN vector or pSIN-LMP1-expressing lentiviruses at the MOI indicated for 5 days. The RON transcripts were detected by RT-Q-PCR and the relative fold increase was normalized to the amounts of RON transcripts in pSIN lentivirus-infected cells (top panel). The protein expression of RON, LMP1, and β-actin was measured by Western blotting (bottom panel). (D) LCLs (2 × 105) were infected with shLuciferase or shLMP1 lentiviruses at an MOI of 4 for 5 days. The RON transcripts were detected by RT-Q-PCR and the relative fold expression of RON was normalized to the amounts of RON transcripts in shLuciferase lentivirus-infected cells (top panel). The protein expression of RON, LMP1 and GAPDH was measured by Western blotting (bottom panel).