Fig 2: Naringenin suppresses ITGA11-associated FAK/PI3K/AKT/mTOR signaling and malignant phenotypes in GC cells. (A) Molecular docking binding mode between Naringenin and ITGA11. (B,C) DARTS assays and quantitative analysis of Naringenin and ITGA11 (n = 3). (D,E) CETSAs and quantitative analysis of Naringenin and ITGA11 (n = 3). (F,G) Cell viability assay of Naringenin in AGS (F) and HGC-27 (G) cells for 24 h (n = 3). (H) Gene expression of ITGA11 following ITGA11 overexpression and Naringenin treatment (20, 40, 80, and 160 µM for 24 h) in AGS cells (n = 4). (I) Representative immunoblot images of ITGA11 and the FAK/PI3K/AKT/mTOR axis following ITGA11 overexpression and Naringenin treatment (20, 40, 80, and 160 µM for 24 h) in AGS cells. (J) Quantification of ITGA11 and FAK/PI3K/AKT/mTOR axis following ITGA11 overexpression and Naringenin treatment (20, 40, 80, and 160 µM for 24 h) in AGS cells (n = 3). (K) Representative images of wound healing, transwell and colony formation assays following ITGA11 overexpression and Naringenin treatment (40 µM; 24 h) in AGS cells. (L) CCK-8 assay following ITGA11 overexpression and Naringenin treatment (40 µM; 24 h) in AGS cells. (M) Quantification of functional assays following ITGA11 overexpression and Naringenin treatment (40 µM; 24 h) in AGS cells (n = 3). (N,O) IF staining of ITGA11 following ITGA11 overexpression and Naringenin treatment (40 µM; 24 h) in HGC-27 cells (n = 3). Scale bars: 50 μm for IF images, 100 μm for transwell images and 500 μm for wound healing images. Data are presented as mean ± s.d. Multiple groups were analyzed by one-way ANOVA. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The original Western blot figures can be found in Supplementary File S1.

Fig 3: ITGA11 activates the FAK/PI3K/AKT/mTOR axis in AGS cells. (A) Representative immunoblot images of the FAK/PI3K/AKT/mTOR axis following ITGA11 overexpression in AGS cells. (B) Quantification of the FAK/PI3K/AKT/mTOR axis following ITGA11 overexpression in AGS cells. (C) Representative immunoblot images of the FAK/PI3K/AKT/mTOR axis following ITGA11 knockdown in AGS cells. (D) Quantification of the FAK/PI3K/AKT/mTOR axis following ITGA11 knockdown in AGS cells. (E) Representative immunoblot images of ITGA11 and the FAK/PI3K/AKT/mTOR axis following ITGA11 overexpression and FAK knockdown in AGS cells. (F) Quantification of ITGA11 and the FAK/PI3K/AKT/mTOR axis following ITGA11 overexpression and FAK knockdown in AGS cells. (G) Representative images of wound healing, transwell and colony formation assays following ITGA11 overexpression and FAK knockdown in AGS cells. (H) CCK-8 assay following ITGA11 overexpression and FAK knockdown in AGS cells. (I) Quantification of functional assays following ITGA11 overexpression and FAK knockdown in AGS cells. Scale bars: 100 μm for transwell images and 500 μm for wound healing images. Data are presented as mean ± s.d. (n = 3 per group). Two-group comparisons were performed using Student’s t test, and multiple groups were analyzed by one-way ANOVA. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The original Western blot figures can be found in Supplementary File S1.

Fig 4: Targeting ITGA11 suppresses gastric cancer progression in vivo. (A) Subcutaneous tumors of ITGA11 overexpression and Naringenin treatment (n = 4). (B) Subcutaneous tumors of ITGA11 knockdown (n = 4). (C,D) Tumor volume (C) and weight (D) of ITGA11 overexpression and Naringenin treatment (n = 4). (E,F) Tumor volume (E) and weight (F) of ITGA11 knockdown (n = 4). (G,H) Representative images of H&E and Ki-67 staining for subcutaneous tumors following ITGA11 overexpression and Naringenin treatment (G) and ITGA11 knockdown (H). (I–K) Gene (I) and protein (J,K) expression of ITGA11 following ITGA11 overexpression and Naringenin treatment in subcutaneous tumors. (L–N) Gene (L) and protein (M,N) expression of ITGA11 following ITGA11 knockdown in subcutaneous tumors (n = 3). Scale bars: 50 μm for H&E and Ki-67 staining. Data are presented as mean ± SEM for tumor volume curves and mean ± s.d. for other analyses. Two-group comparisons were performed using Student’s t test, and panels (C,E) were performed using repeated-measures ANOVA. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001 The original Western blot figures can be found in Supplementary File S1.

Fig 5: Upregulation of ITGA11 in gastric cancer predicts poor prognosis and promotes malignant progression in AGS cells. (A) Representative IHC staining of ITGA11 in paired GC and adjacent normal tissues (Stage II). (B) Quantification of IHC staining scores for ITGA11 from 60 paired GC and normal tissues (n = 60). (C) Kaplan–Meier survival analysis of overall survival in 60 GC patients stratified by ITGA11 expression level. (D–F) ITGA11 mRNA (D) and protein (E,F) expression in 5 pairs of GC and adjacent normal tissues (n = 5). (G) Representative IF staining of ITGA11 in 10 paired GC and adjacent normal tissues (Stage III). (H) Quantification of ITGA11 fluorescence intensity in 10 paired GC and normal tissues (n = 10). (I–K) ITGA11 mRNA (I) and protein (J,K) expression in a panel of GC cell lines (MGC-803, HGC-27, SGC-7901, AGS) relative to the normal gastric epithelial cell line GES-1 (n = 4 for panel (I), and n = 3 for panels (J,K)). (L–N) Gene (L) and protein (M,N) expression of ITGA11 following ITGA11 overexpression (oeITGA11) in AGS cells (n = 6 for panel (L), and n = 3 for panels (M,N)). (O–Q) Gene (O) and protein (P,Q) expression of ITGA11 following ITGA11 knockdown (shITGA11-1/2) in AGS cells (n = 4 for panel (O), and n = 3 for panels (P,Q)). (R,S) CCK-8 assay following ITGA11 overexpression (R) or knockdown (S) in AGS cells (n = 3). (T,U) Wound healing, transwell and colony formation assays following ITGA11 overexpression in AGS cells (n = 3). (V,W) Wound healing, transwell and colony formation assays following ITGA11 knockdown in AGS cells (n = 3). Scale bars: 100 μm for IHC, IF, transwell images and 500 μm for wound healing images. Data are presented as mean ± s.d. Two-group comparisons were performed using Student’s t test, and multiple groups were analyzed by one-way ANOVA. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. The original Western blot figures can be found in Supplementary File S1.