Figure 3.

Activation with ACh enhances formation of the multiprotein complex including SLK, Plk1, and paxillin. (A) Extracts of unstimulated and ACh (10⁻⁴ M, 5 min)-stimulated HASM cells were immunoprecipitated with Plk1 antibody. Blots of the immunoprecipitates and input were probed with antibodies against SLK, Plk1, and paxillin. Protein ratios of SLK/Plk1 and paxillin/SLK in stimulated cells are normalized to the ratios obtained from unstimulated cells. Data are mean values of experiments from four batches of cell culture from three donors. Error bars represent SD. (B) Reverse co-IP analysis also demonstrates that ACh stimulation increases the assembly of the multiprotien complex. Protein ratios of paxillin/SLK and Plk1/SLK in ACh-treated cells are normalized to untreated cells. Data are mean values of experiments from four batches of cell culture from three donors. Error bars indicate SD. *P < 0.05 and **P < 0.01. Student’s t-test was used for statistical analysis.