FIG. 3.

Detection of LMP-1 by Western blotting in CEM and BJA-B cells. (A) Whole-cell lysates (40 μg) from LMP-expressing and control vector-transfected cells were resolved by SDS-10% PAGE. After transfer onto nylon membranes, LMP-1 was detected by anti-LMP monoclonal antibody S12 followed by detection with alkaline phosphatase-conjugated anti-mouse antibodies 5-bromo-4-chloro-3-indolylphosphate (BCIP) and nitroblue tetrazolium. Lanes: 1, BJA-B transfected with the control vector; 2 to 4, three different LMP-expressing BJA-B clones; 5, CEM transfected with the control vector; 6 to 8, three different LMP-expressing CEM clones; 9, an EBV-transformed B-cell line (LCL). The LMP-expressing BJA-B and CEM cells were fractionated into cytoplasmic and nuclear lysates, and each fraction was analyzed for LMP-1 expression by Western blotting. Panels B and C show LMP-1 expression in cytoplasmic and nuclear fractions, respectively. The lanes represent the clones indicated for panel A. The arrows point to LMP-1 bands.