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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. 5 — A The RNA-seq of ASAP1-depletedGC cell and a volcano plot showing the differential genes. B KEGG pathway analysis showing enrichment of differential gene. C The whole cell lysates were extracted from SGC-7901 cells expressing ASAP1-HA or from ASAP1^+/− SG-7901 cells to measure CDC42, RAC1, RALA and RhoA GTPase activity by G-LISA activation assay. D The whole cell lysates prepared from either SGC-7901 or MGC803 cell expressing ASAP1-HA were used for CDC42 activation assay followed by western blotting. E Combined phalloidin staining for F-actin (red) and ASAP1 (green) immunofluorescence. Nuclei were counterstained with DAPI (blue). In ASAP1^+/− SGC-7901 cells, the filamentous pseudopodia disappeared and cytoskeleton depolymerized. F SGC-7901 cells were treated with CDC42 inhibitor ZCL278 and combined phalloidin staining for F-actin (red). The filamentous pseudopodia disappeared and cytoskeleton depolymerized. G SGC-7901 cells were treated with CDC42 inhibitor ZCL278 and the whole cell lysates were extracted to measure CDC42 GTPase activity by G-LISA activation assay. H The proliferation of SGC-7901 cells transfected with pcDNA3.1, SGC-7901 cells transfected with pcDNA3.1-ASAP1-HA, SGC-7901 cells treated with ZCL278 or SGC-7901 cells transfected with pcDNA3.1-ASAP1-HA followed by treated with ZCL278 was determined by CCK-8. I The migration of various types of cells described in H was determined by Transwell assays.