Figure 5
Figure 5. LMP2A inhibited the E47 and PU.1 promoter activity. (A) Schematic map of the promoter region of PU.1 (−470 to +57). Deletion constructs derived from the PU.1 construct (−470 to +57) were subcloned into the pCDH-GL3 luciferase reporter vector. BJAB cells were infected with pSIN- and LMP2A-expressing lentiviruses. After 3 days, the cells were infected with the reporter lentiviruses indicated. After 4 days, luciferase activities from each transfectant were normalized with the GFP intensities. (B) BJAB cells were transduced with pLKO or PU.1 lentiviruses. After 5 days, infected BJAB cells were transduced with PU.1- (−470 to +57), pSIN-, or LMP2A-expressing lentiviruses and then the luciferase activities were measured and normalized to GFP activity at day 5 postinfection. (C) BJAB cells were transduced with shLuc or shPAX5 lentiviruses. After 5 days, infected BJAB cells were further transduced with PU.1- (−470 to +57), pSIN-, or LMP2A-expressing lentiviruses. At day 5 postinfection, the luciferase activities were measured and normalized to GFP activity. (D) Schematic map of the promoter region of E47 (−1000 to +29). Deletion constructs derived from the E47 construct (−1000 to +29) were subcloned into the pCDH-GL3 luciferase reporter vector. The 293T cells were transfected with pSG5- and LMP2A-expressing plasmids. After 3 days, the luciferase activities were measured and normalized to GFP activity. (E) The 293T cells were transfected with pCDH-GL3-E47– (−1000 to +29), pSG5-, or LMP2A-expressing plasmids and treated with 500 nM Mithramycin. At day 3 postinfection, the luciferase activities were measured and normalized to GFP activity. (F) The 293T cells were infected with shLuc or shSP1 lentiviruses. After 5 days, infected 293T cells were transfected with pCDH-GL3-E47– (−1000/+29), pSG5-, or LMP2A-expressing plasmids. At day 3 postinfection, the luciferase activities were measured and normalized to GFP activity. Each experiment was performed 3 times, and 1 representative is shown.

LMP2A inhibited the E47 and PU.1 promoter activity. (A) Schematic map of the promoter region of PU.1 (−470 to +57). Deletion constructs derived from the PU.1 construct (−470 to +57) were subcloned into the pCDH-GL3 luciferase reporter vector. BJAB cells were infected with pSIN- and LMP2A-expressing lentiviruses. After 3 days, the cells were infected with the reporter lentiviruses indicated. After 4 days, luciferase activities from each transfectant were normalized with the GFP intensities. (B) BJAB cells were transduced with pLKO or PU.1 lentiviruses. After 5 days, infected BJAB cells were transduced with PU.1- (−470 to +57), pSIN-, or LMP2A-expressing lentiviruses and then the luciferase activities were measured and normalized to GFP activity at day 5 postinfection. (C) BJAB cells were transduced with shLuc or shPAX5 lentiviruses. After 5 days, infected BJAB cells were further transduced with PU.1- (−470 to +57), pSIN-, or LMP2A-expressing lentiviruses. At day 5 postinfection, the luciferase activities were measured and normalized to GFP activity. (D) Schematic map of the promoter region of E47 (−1000 to +29). Deletion constructs derived from the E47 construct (−1000 to +29) were subcloned into the pCDH-GL3 luciferase reporter vector. The 293T cells were transfected with pSG5- and LMP2A-expressing plasmids. After 3 days, the luciferase activities were measured and normalized to GFP activity. (E) The 293T cells were transfected with pCDH-GL3-E47– (−1000 to +29), pSG5-, or LMP2A-expressing plasmids and treated with 500 nM Mithramycin. At day 3 postinfection, the luciferase activities were measured and normalized to GFP activity. (F) The 293T cells were infected with shLuc or shSP1 lentiviruses. After 5 days, infected 293T cells were transfected with pCDH-GL3-E47– (−1000/+29), pSG5-, or LMP2A-expressing plasmids. At day 3 postinfection, the luciferase activities were measured and normalized to GFP activity. Each experiment was performed 3 times, and 1 representative is shown.

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