Fig. 6

Most DLC1 START mutants show reduced binding to PS and PS supplementation does not improve their binding to Caveolin-1 or PLCD1. A Pull-down (PD) using PS-coated beads in 293 T cells overexpressing GFP or GFP DLC1 wild type (WT) or mutants followed by GFP western blot (WB). Expression of GFP fusion proteins was checked by western blot in total lysates (bottom panel). B, C Immunoprecipitation (IP) using PLCD1 antibody (top) or Caveolin-1 antibody (bottom) in 293 T cells overexpressing GFP DLC1 wild type (WT) or mutants in the presence or absence of 100 μM PS added to the binding reaction, followed by Western blot (WB) using the indicated antibodies. The expression of GFP fusion proteins was checked by Western blot in total lysates (bottom panel)