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. 2012 Sep 3;198(5):865–880. doi: 10.1083/jcb.201204107

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© 2012 Bastos et al.

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Figure 4. — RhoA and Rac1 couple to discrete effector pathways in mitotic cells. (A) Effector binding assays from mitotic HeLa cell lysate were performed using GST, RhoA, Rac1, and Cdc42 as described in the Materials and methods. Bound fractions were eluted using sample buffer and analyzed by SDS-PAGE and MS. Coomassie brilliant blue–stained gels of the RhoA, Rac1, and Cdc42 complexes are shown. Proteins identified by MS are listed in the table and marked by the side of the appropriate gel. Peptide number and intensity give a measure of the relative abundance. (B) Western blot analysis of the input and bound effector proteins was performed using the antibodies shown in the figure. (C) Effector binding assays from interphase (Int) and mitotic (Mit) HeLa cell lysate were performed using GST, RhoA, and Rac1 as described in the Materials and methods. Samples of the input material and bound fractions were analyzed by Western blotting using the antibodies shown in the figure.