Fig. 4.

E2F-1 associated pRB is O-GlcNAc modified. E2F-1 was immunopurified from 3T3-L1 preadipocytes with an antibody to E2F-1, as well as normal rabbit IgG as a negative control, in early G1 (4 h) of the cell-cycle, and the resulting proteins separated by SDS–PAGE. Following transfer, the presence of co-purifying pRB was detected by Western blotting and O-GlcNAc modified pRB was detected by lectin blotting with sWGA coupled to HRP