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. 2003 Apr;77(8):5008–5013. doi: 10.1128/JVI.77.8.5008-5013.2003

FIG. 4.

FIG. 4.

Activation of LMP1 promoter by combinations of EBNA2 and SMN mutants (A) Coactivation of the LMP1 promoter by SMN and EBNA2ΔRG. The EBNA2wt and EBNA2ΔRG expression vectors were transfected with or without the SMNwt expression plasmid and tested for activation of the LMP1 promoter (−327/+40) in BJAB cells as described in the work of Voss et al. (30). (B) Expression of HA-tagged SMNwt and an HA-tagged SMN expression construct with a deletion of the Tudor domain (SMNΔTudor) in HeLa cells after transient transfection, separation by SDS-10% PAGE, and detection with HA-specific MAb 3F10. Numbers at left are molecular masses in kilodaltons. (C) SMNΔTudor with a deletion of the Tudor domain fails to coactivate the LMP1 promoter. The SMNΔTudor mutant was expressed with EBNA2wt or EBNA2ΔRG in BJAB cells and tested for activation of the LMP1 promoter construct. (D) No significant coactivation of the LMP1 promoter by EBNA2 and an SMA patient-derived point mutant in the Tudor domain of SMN (SMN E134K). The EBNA2wt expression vector was transfected with SMNwt or SMN E134K and tested for activation of the LMP1 promoter in BJAB cells. The experiments the results of which are shown in panels A and C were carried out five times each in duplicate; the experiments the results of which are shown in panel D were carried out three times in duplicate.