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Fig. 5 | Molecular Medicine

Fig. 5

From: Lasp-1, a Novel Type of Actin-Binding Protein Accumulating in Cell Membrane Extensions

Fig. 5

Actin binding of Lasp-1. (A) Schematic representation of mouse GST-Lasp, GST-N-Lasp, and GST-C-Lasp. Symbols are as in legend of Figure 1A. GST is represented by a cross-hatched box. (B) Mouse Lasp-1 interacts with endogenous actin in COS-1 cells. GST-Lasp, GST-N-Lasp, and GST-C-Lasp (lanes 2 and 2′, 3 and 3′, and 4 and 4′ respectively) expressed in COS-1 cells, and purified by GST-pull down were separated by SDS-PAGE and transferred to nitrocellulose. The blot was first probed with the AC-40 anti-actin antibody (right panel), stripped, then reprobed with an anti-GST antibody (left panel). Lanes 1 and 1′: COS-1 cell lysate. (C) Farwestern experiment showing the binding of mouse Lasp-1 to purified actin. GST-Lasp, GST-N-Lasp, and GST-C-Lasp (lanes 2 and 2′, 3 and 3′, and 4 and 4′ respectively) immobilized on nitrocellulose sheets as in (B) were incubated 2 hr with purified actin (25 µg/ml) before immunodetection with the AC-40 anti-actin antibody (right panel). The same blot was reprobed with anti-GST antibody (left panel). Lanes 1 and 1′: COS-1 cell lysate.

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