Figure 6
Functional regions of EWS-Oct-4B. (A) Transcriptional properties of EWS-Oct-4B deletion mutants. Shown on the left are schematic representations of EWS-Oct-4B deletion constructs. A reporter plasmid, pOct-4(10 × ) TATA luc, was cotransfected into 293T cells with various flag-tagged EWS-Oct-4B mutants. Relative transcriptional activation values are shown on the right as mean increases±s.e. relative to a value of 100% for transfection of EWS-Oct-4B. The results are the means of three independent experiments performed in duplicate. (B) Immunoblot analysis showing expression of the EWS-Oct-4B deletion mutants in transiently transfected cells. Total cell lysates were fractionated by 12% SDS–PAGE and visualised by western blotting with anti-Flag (M2, Sigma, top panel) or anti-EGFP (Invitrogen Molecular Probes, bottom panel) antibodies. The positions of pre-stained molecular weight markers (New England Biolabs, Hitchin, UK) are indicated to the left (kDa). Lane 1, empty vector; lane 2, EWS-Oct-4B; lane 3, EWS-Oct-4B (ΔEWS); lane 4, EWS-Oct-4 (ΔCTD); and lane 5, EWS-Oct-4B (V313P).
