FIGURE 1.

Endogenous hCLE complexes bind cap analog resins. (A) Nuclear and cytoplasmic extracts of HEK293T cells were incubated with control (4B) or cap analog (cap) resins. Western blot analysis of bound proteins using the corresponding antibodies are shown. (B) Graph showing the percentage of the indicated proteins bound to m⁷GTP analog resin (blue) or control resin (yellow), compared to the initial input that was incubated with the corresponding resins. Total extracts and the data of three independent experiments were used. (C) Total protein extracts from HEK293T cells were incubated with cap analog resin and eluted with 1 mM m⁷GTP. Eluted fractions and resin-bound proteins were resolved in non-denaturing gels and analyzed by Western blot with indicated antibodies. ^∗monomer, ^∗∗dimers, and ^∗∗∗trimers. Cap-binding experiments were repeated more than five times. Input:bound protein ratios were: 1:12 in total and cytoplasmic HEK293T cell extracts; 1:6 in HEK293T cell nuclear extracts.