Skip to main content
Fig. 4 | Journal of Hematology & Oncology

Fig. 4

From: MIIP remodels Rac1-mediated cytoskeleton structure in suppression of endometrial cancer metastasis

Fig. 4

MIIP blocking of cell migration through competing with Rac1-GTP to bind the PAK1-binding domain. a Competition for PAK1 PBD binding between MIIP and Rac1. HEC1B cell lysate with MIIP loaded with GDP (different amounts of MIIP) and control HEC1B lysate loaded with GTPγS (same amounts of Rac1/GTP) were incubated with GST-PAK1 PBD-coupled agarose beads, and the pulled down complex was detected by indicated antibodies. The original cell lysate loaded with GTPγS or GDP was used as positive or negative control, respectively. b In an immunoprecipitation assay using anti-MIIP antibody, PAK1 was pulled down by MIIP. c In a reverse immunoprecipitation assay using anti-PAK1 antibody, MIIP was pulled down by PAK1. Asterisk indicates IgG heavy chain. d Co-localization of MIIP and PAK1 in HEC1B cells. HEC1B cells were transfected with HA-tagged MIIP for 48 h and then stained with HA (green) and PAK1 (red). All images were taken by ZEISS HEO 100 microscope at a magnification of ×630. Scale bar 10 μm. Arrow: lamellipodia. Arrow head: MIIP and PAK1 co-localization. e Immunoprecipitation assays used rabbit anti-HA in HEC1B cells transfected with wild-type MIIP or one of the four truncated MIIP constructs. Input: immunoblot of a steady level of MIIP, Rac1, or PAK1 in HEC1B cell lysates (30 % of the amount of the same cell lysate sample used for immunoprecipitation). IgG H indicates IgG heavy chain. C, 1, 2, 3, 4, and 5 indicate control, wild-type MIIP, MIIP (155-388), MIIP (1-360), MIIP (1-313), and MIIP (1-283), respectively. f, g The effects of different truncated MIIPs on cell migration. f Representative photographs of different truncated MIIPs on cell migration by transwell migration assay (magnification ×200). g Data are expressed as means ± SD of cells per 10 high-power fields (magnification ×200) from three separate experiments

Back to article page