Figure 4
Production of a super-shifted H3 band in RKO and patient tissue extracts by super-shift EMSA with a monoclonal antibody against U2AF65. 33P-labeled triplex DNA (1 nM) was complexed with 1.5 μg total protein from RKO cytoplasmic (lanes 2-4), RKO nuclear (lanes 5-7), 5 μg tumor cytoplasm (T cyto lanes 8-10) or tumor nuclear (T nuc lanes 11-12) extracts. Lanes 2, 5, 8, and 11, no antibody; lanes 3, 6, 9, and 12, 400 ng anti-U2AF65 antibody MC3; lanes 4, 7, and 10, mouse IgG antibody (negative control). Each reaction also contained 2 μg poly (dI-dC) carrier DNA. Lane 1, triplex DNA probe alone.