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. 2008 Jun 2;181(5):747–760. doi: 10.1083/jcb.200710111

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© 2008 Mondal et al.

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Figure 8. — RasGEF Q regulates myosin II through MHCK A. (A) GST-GEF can associate with myosin II and MHCK A in a manner that is sensitive to Lat A treatment. (B) Cytoskeletal fractions from wild-type AX2, AX2 cells overexpressing GFP-GEF, and gefQ⁻ cells were prepared as described in Materials and methods. Supernatant and pellet fractions were resolved by SDS-PAGE and immunoblotted using MHCK A–specific monoclonal antibody. The graph below represents the mean of two independent experiments and shows the amount of MHCK A present in the pellet fractions as the percentage of the total MHCK A content. (C) Presence of MHCK A in fractions containing active RasB. Active RasB was pulled down from GFP-GEF (6 h) cells using GST-Byr2(RBD). The precipitates were tested for the presence of MHCK A and myosin II heavy chain.