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. 2023 Feb 15;14(2):124. doi: 10.1038/s41419-023-05648-9

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — A The ASAP-associated interactomes were determined by immunoprecipitation and mass spectrometry (IP-MS). Venn diagrams show the number of proteins that may interact listed in the tables. B Correlation analysis of ASAP1 and IQGAP1 RNA expression in TCGA STAD database. C Images showing the immunofluorescence of ASAP1 (red) and IQGAP1 (green). Nuclei were counterstained with DAPI (blue). C SGC-7901 was transfected with pcDNA3.1 or pcDNA3.1-ASAP1-HA. 24 h after transfection, HA-tag fused proteins were immunoprecipitated with their interacting proteins. Western blot was used to detect indicated proteins. D Western blot showing that endogenous ASAP1 co-immunoprecitated with IQGAP1 from SGC-7901 cells. An anti-ASAP1 antibody was used for IP. Normal mouse IgG was used as a control. E Western blot showing that HA-tagged ASAP1 coimmunoprecipitated with IQGAP1 from SGC-7901 cells expressing HA-tagged ASAP1, but not from control SGC-7901 cells. An anti-HA antibody was used for IP. F Molecular docking pattern diagram of ASAP1 and IQGAP1. G IQGAP1 mRNA level of MGC-803 and SGC-7901 after ASAP1 overexpression. H IQGAP1 protein level of MGC-803 and SGC-7901 after ASAP1 overexpression. I SGC-7901 and SGC-ASAP1^+/− cells were incubated with MG132 (+) or DMSO (−) for 4 h. Equal amounts of protein lysates were loaded directly onto gels or immunoprecipitated (IP) with an anti-IQGAP1 antibody. Proteins were analyzed by SDS–PAGE and immunoblotting (IB) using anti-ubiquitin and anti-IQGAP1 antibodies, respectively. J Western blot showing that IQGAP was depleted by two different siRNA oligos in SGC-7901 cells. K The proliferation of various types of SGC-7901 cells was determined via CCK-8, including cells transfected with pcDNA3.1, cells transfected with pcDNA3.1-ASAP1-HA, cells transfected with pcDNA3,1 followed by transfection with a siRNA oligo against IQGAP1, and cells transfected with pcDNA3.1-ASAP1-HA followed by transfection with a siRNA oligo against IQGAP1. M The migration of various types of SGC-7901 cells described in L was determined was determined by transwell assays. N The who cell lysates were extracted from various types of SGC-7901 cells to measure CDC42 GTPase activity by G-LISA activation assay, including cells transfected with pcDNA3.1, cells transfected with pcDNA3.1-ASAP1-HA, and cells transfected with pcDNA3.1-ASAP1-HA followed by transfection with a siRNA oligo against IQGAP1.