FIG. 5.

BS69 is required for the complex formation between LMP1 and TRAF6. (A) The empty shRNA vector (pSuper) or different BS69 shRNA constructs were transfected to 293T cells twice, followed by transfection of an expression vector encoding HA-BS69. WCEs were subjected to immunoblotting with the anti-HA antibody. MT, mock transfected; N2, an ineffective BS69 shRNA clone with 2-bp mutations in its sequence; N3, the correct BS69 shRNA construct targeting a region in the amino terminus of BS69; C2 and C3, two correct clones of BS69 shRNA targeting a region in the carboxyl terminus of BS69. (B) 293T cells were first transfected twice with either pSuper or BS69 shRNA (C3), followed by transfection of an empty vector or LMP1. WCEs were first immunoprecipitated with the anti-LMP1 antibody; both the immunoprecipitated LMP1 and the coprecipitated TRAF6 were sequentially detected by immunoblotting.