FIGURE 1:

Identification of ROCK1 as a p120 binding partner. (A) Distribution of ROCK1 peptides recovered from p120 ReCLIP eluates described previously (Smith et al., 2011). Small numbers refer to amino acid position of specific domains in the ROCK1 structure. Large numbers refer to the number of unique ROCK1 peptides recovered per domain (see Table 1). (B) Coimmunoprecipitation of ROCK1 with p120. Lane 1: whole-cell lysates containing 10% of input. Lanes 2–4: immunoprecipitates from DSP–cross-linked A431 cells were generated with two separate p120 mAbs (15D2 and pp120) or a control monoclonal antibody (IgG) as indicated, and detected by Western blotting with antibodies to ROCK1 or p120. (C) Coimmunoprecipitation of ROCK1 with p120 in the absence of cross-linker. p120 was immunoprecipitated from DSP–cross-linked (lane 1) and un-cross-linked (lane 3) A431 cells. Lanes 4 and 6: whole-cell lysates from DSP–cross-linked (lane 3) and un-cross-linked (lane 4) A431 cells. Lanes 2 and 5 were left empty to avoid cross-contamination.