Fig. 2: Exon 7 of LRRFIP2 variant 3 determines the metastatic potential of gastric cancer cells in vitro and in vivo.

a Transwell migration assay and Matrigel invasion assay of MKN1 cell lines stably expressing ESRP1 proteins (left) and bar graphs showing number of invaded and migrated cells (right), respectively, following staining with crystal violet. Original magnification, _40X. Scale bar, 0.5 mm. b RT-PCR analysis showing overexpression of ESRP1 and LRRFIP2 variant 2 and 3. c Immunoblot and RT-PCR analysis of ESRP1 immunoprecipitation. d Immunoblot and RT-PCR analysis of LPCX-Flag-LRRFIP2 variants 2 and 3-overexpressing MKN28 cells. e Transwell migration assay and Matrigel invasion assay of MKN28 cell lines stably expressing LRRFIP2 variant 2 and 3 proteins (left) and bar graphs showing number of invaded and migrated cells (right), respectively, following staining with crystal violet. Original magnification, _40X. Scale bar, 0.5 mm. f Representative whole liver image showing metastatic nodules (left) and scatter plot showing the number of liver metastatic nodules (right). g H&E staining showing sections of the metastasized liver from f. Original magnification, _100X. Scale bar, 100 µm. h Schematic representation of exon 7 deletion by CRISPR/Cas9 system. i RT-PCR analysis of LRRFIP2 in exon 7-deleted cell lines. j Transwell migration assay and Matrigel invasion assay of exon 7-deleted cell lines (top) and bar graphs showing number of invaded and migrated cells (bottom), respectively, following staining with crystal violet. Original magnification, _40X. Scale bar, 0.5 mm. k Representative whole liver image showing metastatic nodules (left) and scatter plot showing the number of liver metastatic nodules (right). l H&E staining showing sections of the metastasized liver from k. Original magnification, _100X. Scale bar, 100 µm. a, e, j Data are representative mean ± SD of three independent experiments (N = 3). b–d, i The representative results were obtained from at least three independent experiments. f Data are representative mean ± SD of five independent animals (n = 5). f Data are representative mean ± SD of four independent animals (n = 4). All P values were calculated by unpaired two-tailed Student’s t tests. These data represent the mean ± S.D. Source data are provided in the Source Data file.