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Figure 2


Fig. 2. PRMT1 colocalizes and interacts with hCAF1. (A) Laser-scanning confocal microscopy was used to compare the distributions of (a) hCAF1 and (b) PRMT1 in MCF-7 cells using the polyclonal anti-hCAF1 and the monoclonal anti-PRMT1 antibodies. The corresponding overlay is shown in panel c and the corresponding phase-contrast micrograph is shown in panel d. (B) MCF-7 cell extract was immunoprecipitated using anti-hCAF1 polyclonal antibody or non-immune serum (NIS). Total cell lysate (input) as well as immunoprecipitants (IP) were analyzed by western blotting using the polyclonal anti-PRMT1 or anti-hCAF1 antibodies. Input represents 4% of the total cell extract used for immunoprecipitation experiments. (C) Direct interaction between hCAF1 and PRMT1 was analyzed by GST-pull-down experiments. 35S-labeled in vitro translated hCAF1, BTG1 and luciferase (left) or deletion mutants (right) were incubated with GST-PRMT1–glutathione-Sepharose beads. The eluted proteins and 1/50 of input radiolabeled proteins were analyzed by SDS-PAGE and visualized by autoradiography.





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