Fig. 5

ANGPTL4 knockdown attenuates the oncogenic effect of D-2HG in TNBC cells via integrin-regulated pathway. A Representative images and quantification of the EdU assay in TNBC cells. Scale bar: 100 μm. B Representative images and quantification of migration and invasion assays in MDA-MB-231 cells. Scale bar: 200 μm. C Representative images and quantification of migration and invasion assays in MDA-MB-468 cells. Scale bar: 200 μm. D TNBC cells were treated with D-2HG for the indicated concentration, and activation of the integrin/SRC/FAK/JAK2/STAT3 pathway was evaluated by western blot. E Western blot analysis of integrin/SRC/FAK/JAK2/STAT3 pathway expression in ANPGTL4 knockdown TNBC cells following D-2HG treatment (500 μM) for 48 h. F Western blot analysis of expression of proteins involved in cell cycle, migration, and invasion in ANPGTL4 knockdown TNBC cells following D-2HG treatment (500 μM) for 48 h. G TNBC cells were pre-treated with 5 ng/mL GLPG0187 for 12 h, followed by D-2HG treatment (500 μM) for 48 h, western blot showed the protein expression related to the integrin pathway. H Images (left) and quantification (right) of lung metastatic nodules from BALB/c nude mice at the endpoint (n = 5/group). I BLI images and quantification (right) of BALB/c nude mice intravenously injected with Angptl4 knockdown 4T1 cells followed by administration of D-2HG or vehicle (n = 5/group). *P < 0.05, **P < 0.01, ***P < 0.001