FIG. 5.

The oncogenic fusion protein EWS–FLI-1 does not coimmunoprecipitate with the TFIID complex in Ewing sarcoma cell lines. (A) The PAb raised against the N-terminal domain of EWS recognizes both wild-type EWS and the two different EWS–FLI-1 fusion proteins in NEs from the two Ewing sarcoma cell lines, RD-ES (lane 2) and COH (lane 3). NEs from HeLa, RD-ES and COH cells were analyzed by Western blotting using the anti-EWS (α-EWS) antibody (upper panel), the anti-FLI-1 (α-FLI-1) MAb (middle panel), and the anti-TBP (α-TBP) MAb 3G3 (lower panel). M, markers in kilodaltons. (B) NEs from the various cell lines were immunoprecipitated (IP) with the anti-TBP MAb 3G3 (lane 1 to 3). Beads were washed and boiled, and bound proteins were analyzed by Western blotting with the anti-EWS antibody and the anti-TBP MAb. The control immunoprecipitation using an unrelated MAb is shown in lane 4.