Fig. 6: EZM2302 reduces metastatic potential of gastric cancer cells by attenuating SERPINE1 expression in the presence of LRRFIP2 variant 3.

a Immunoblot analysis showing expression of H3R17me2a and histone H3 in MKN1 cells upon EZM2302 treatment for indicated hours. b Immunoblot analysis showing expression of H3R17me2a, histone H3, and Flag in MKN28 cells overexpressing LRRFIP2 variant 3 upon EZM2302 treatment for indicated hours. c, d qChIP analysis of the human SERPINE1 promoter upon treatment of EZM2302 for 24 h with antibodies to CARM1, H3R17me2a in c LRRFIP2 variant 2 and 3-overexpressing MKN28 cell lines, d LRRFIP2 variant 3 exon7 deleted MKN1 cell lines. e Real-time qRT-PCR analysis showing SERPINE1 expression in MKN1 cells upon EZM2302 treatment for indicated hours. f RT-PCR analysis showing SERPINE1 expression in MKN28 cells overexpressing LRRFIP2 variant 3 upon EZM2302 treatment for indicated hours. g, h Transwell migration assay and Matrigel invasion assay of g MKN28 cells overexpressing LRRFIP2 variant 2 and 3 and h LRRFIP2 variant 3 exon 7 deleted MKN1 cells with EZM2302 treatment (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. P values verses Control 0 µM, otherwise indicated. i Representative whole liver image showing metastatic nodules (left) and scatter plot showing the number of liver metastatic nodules (right). j Representative IHC images showing SERPINE1 and H3R17me2a expression and H&E staining in metastasized liver tissues from i (left) and H-score scatter plots of the IHC staining (right). Original magnification, _200X. Scale bar, 50 µm. a, b The representative results were obtained from at least three independent experiments. c, d Data are representative mean ± SD of three independent samples (N = 3). g, h Data are representative mean ± SD of three independent experiments (N = 3). i Data are representative mean ± SD of five independent animals (n = 5). j Data are representative mean ± SD of six independent measurements (n = 6). All P values were calculated by unpaired two-tailed Student’s t tests. n.s: not significant. Source data are provided in the Source Data file.