Fig 2: Overexpression of TMEM100 inhibits the cell cycle and apoptosis of PCa cells and regulate FAK/PI3K/AKT signaling pathway. (A) Effects of TMEM100 overexpression on cell cycle by flow cytometry in DU145 and PC3 cells. (B) Effects of TMEM100 overexpression on cell apoptosis by flow cytometry in DU145 and PC3 cells. (C) Changes in the apoptotic and cell cycle-related proteins were detected by Western blot in DU145 and PC3 cells. (D) The analysis results of enrichment of KEEG pathway. (E) Changes in the FAK/PI3K/AKT signaling pathway proteins were detected by Western blot in DU145 and PC3 cells. (F) The qualification of apoptotic and cell cycle-related proteins by densitometry (G) The qualification of FAK/PI3K/AKT signaling pathway proteins by densitometry. *P<0.05 VS control group.

Fig 3: TMEM100 inhibits migration, invasion and EMT by regulating SCNN1D through FAK/PI3K/AKT signaling pathway. (A-B) The depletion of SCNN1D increases migration and invasion ability in TMEM100-overexpression DU145 and PC3 cells. (C) The depletion of SCNN1D rescue the function that TMEM100 overexpression hindered the protein level of p-FAK, p-PI3K and p-AKT. (D) TMEM100 affected cell apoptosis, cell cycle and EMT via regulating SCNN1D by western blot analysis. *P<0.05 VS control group, #P<0.05 VS TMEM100 overexpression group.

Fig 4: MYLK-AS1/miR-424-5p/E2F7 axis positively regulates HCC metastasis, invasion, and angiogenesis via the VEGFR-2 signaling pathway. a E2F7, VEGFR-2, p-VEGFR-2, PLC-λ, p-PLC-λ, Erk1/2, p-Erk1/2, P38, p-P38, FAK, p-FAK, Src, p-Src, Akt, and p-Akt protein expression in Hep-G2 or MHCC-97H cells divided in the following groups: Control, shRNA-NC, shMYLK-AS1-2, NC, and MYLK-AS1. NC represents a blank plasmid control, and MYLK-AS1 is a group overexpressing this lncRNA. The numbers represent the quantification of the relative protein amount. GAPDH was used as the loading control. VEGFR-2 and p-VEGFR-2 protein expression detected after 7-day-co-culture of HUVECs with Hep-G2 or MHCC-97H cells divided in the same groups as above. GAPDH was used as the loading control. b Schematic diagram of the regulatory mechanism of MYLK-AS1/miR-424-5p/E2F7 axis in the promotion of HCC proliferation, metastasis and angiogenesis

Fig 5: Identification of ITGA2-dependent activation of FAK and MAPK signaling axis using phosphoproteomics.(A) Venn-diagram highlights the number of significantly downregulated phosphoproteins shared among three ?ITGA2 ovarian cancer cell lines. (B) Top seven gene ontology (GO) biological process terms with highest statistical significance. The horizontal axis displays the number of genes in the intersection group of three ?ITGA2 ovarian cancer cell lines. (C) GSEA analysis identified up- or downregulated pathways in ?ITGA2 cells. The Cluster Profiler dot plot visualization shows enriched terms as dots. The highest ranking of the significantly enriched pathway is displayed. (D) Reactome pathway analysis of MAPK and cell junction organization pathway comparing ?ITGA2 with WT cells. NES = normalized enrichment score. Nominal p-value calculated by the permutation test in GSEA analysis. (E) Volcano plot shows differentially expressed phosphoproteins between WT and ?ITGA2 cells. y-axis defines the statistical significance -Log10p<2 and x-axis defines the magnitude of Log2-fold change >2 or < -2. Colored circle defines significantly decreased (blue) or increased (red) phosphopeptides in ?ITGA2 cells. (F) Heatmap analysis of KEGG hsa04510:focal adhesion pathway phosphoproteins. Data were analyzed based on three biological replicates of IGROV1 WT and ?ITGA2 cells. (G) Western blot analysis shows major change of phosphoproteins in the focal adhesion signaling upon treatment of 5 µM Defactinib for 24 hr. (H) Western blot analysis shows induced activation and phosphorylation of FAK/Src/MAPK signaling in patient-derived ITGA2high compared to ITGA2low omental tumor (n = 10). Figure 7—source data 1.Analysis of phosphoproteomics in IGROV1, SKOV3, and OVCAR3 WT versus ?ITGA2 cell lines.