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. 2008 Apr 7;105(14):5489–5494. doi: 10.1073/pnas.0801053105

Fig. 5.

Fig. 5.

S18-2 binds EBNA-b and pRb in vitro specifically. (A) GST-S18-2 precipitates pRb regardless of EBNA-6 presence in GST pulldown assay. The membranes were probed with anti-pRb (Upper) and anti-ppRb (Lower) antibodies. Lane 1: lysate of 0.4 × 106 MUTUI cells; lane 2: lysate of 0.4 × 106 MUTUIII cells; lane 3: GST precipitation from MUTUI cell lysate (10 × 106 cells); lane 4: GST precipitation from MUTUIII cell lysate (10 × 106 cells); lane 5: GST-S18-2 precipitated pRb and ppRb from the MUTUI cell lysates (10 × 106 cells); lane 6: GST-S18-2 precipitated pRb and ppRb from MUTUIII cell lysates (10 × 106 cells). (B) GST-S18-2 precipitates pRb specifically in GST pulldown assay. The membranes were probed with anti-p130, anti-pRb, and anti-E2F1 antibodies. Lane 1: MUTUIII cell lysate; lane 2: GST beads; lane 3: GST-S18-2 precipitated pRb, but not p130 neither E2F1. (C) GST-S18-2 precipitates EBNA-6 and pRb simultaneously in GST pulldown assay. Membranes were probed with anti-EBNA-6 and pRb antibody. Lanes 1, 3, and 5: RAJI cell lysate, lanes 2, 4, and 6: RAJI-EBNA-6 cell lysate. Lanes 1 and 2: cell lysates (5% of input); lanes 3 and 4: GST beads; lanes 5 and 6: GST-S18-2 beads.