Skip to main content
. 2011 Oct 3;108(41):17129–17134. doi: 10.1073/pnas.1112122108

Fig. 3.

Fig. 3.

The LD mutation but not the GAP dead mutation in full-length DLC1 reduces complex formation and colocalization with endogenous talin. (A) Pull-down assay in transfected 293T cells. The WT DLC1 and the GAP dead DLC1 R718A with or without deletion of LD motif (delLD) were pulled down by GST-talin (1,288–1,646) followed by immunoblotting. The expression of transfected proteins from WCE is shown as loading controls. (B) Complex formation of DLC1 with endogenous talin in stably transfected H358 cells, which do not express detectable endogenous DLC1. Characterization of H358 stable clones expressing R718A with or without delLD: expression, Rho-GTP, and coimmunoprecipitation with endogenous talin. (C) Colocalization of DLC1 with endogenous talin in H358 stable cell lines. Coimmunostaining of endogenous talin (red) and DLC1 (green) in H358 parental cells and stable clones expressing DLC1-R718A with or without delLD mutation. The confocal images are representative of the majority of cells observed. (Scale bar: 10 μm.) (D Upper) Pull down of endogenous talin in 293T cells cotransfected with GST or GST-fusion and DLC1 fragment (448–500) containing WT or mutant LD motif (mtLD) followed by immunoblotting. (D Lower) The conserved WT LD-like motif in DLC1, the three substitution mutations (underlined) in mtLD, and the deleted LD amino acids in delLD are shown.