Fig. 8
From: Structure of SgK223 pseudokinase reveals novel mechanisms of homotypic and heterotypic association
SgK223-SgK269 heterotypic association in cells and SgK223 function. a Co-immunoprecipitation of exogenous HA-tagged version of FL WT SgK223 or mutant proteins with endogenous SgK269 in MCF-10A SgK223 KO cells. Lysates were subjected to immunoprecipitation with an anti-HA antibody followed by western blotting with anti-HA or anti-SgK269 antibodies as indicated. Total cell lysates were also blotted with these antibodies, as well as for actin as a loading control. The data are representative of two independent experiments. b Effect of interface mutations on SgK223-promoted cell migration. Western blotting of total cell lysates to confirm expression levels of SgK223 FL WT and mutants in MCF-10A parental and SgK223 KO cells. Lysates were blotted with the indicated antibodies (top). Effect of SgK223 FL WT and mutants on cell migration in MCF-10A SgK223 KO cells (bottom). Relative wound closure was measured using TScratch software59 and used to calculate the cell migration rate. This is expressed relative to vector control, which was arbitrarily set at 1.0. Data represent mean ± SEM from four independent experiments. ***p < 0.005 (for comparison of WT to vector), *p < 0.05, **p < 0.01 (for comparison of mutants to WT). See also Supplementary Fig. 7
