Fig. 4

 F. nucleatum-derived EVs contribute to BC cell proliferation, migration, and invasion via activation of TLR4 A. The expression of TLR4 protein in BC cells with different treatments was examined by western blot (n = 3 per group) B. The transfection efficiency of sh-TLR4#1, #2, and #3 was determined by (top) RT-qPCR and (bottom) western blot (n = 3 per group). MDA-MB-231 cells were treated with PBS, Fn-EVs, or sh-TLR4 transfection combined with Fn-EVs (n = 3 per group) C. The cell viability was detected by CCK-8 after diverse treatments for 24, 48, and 72 h D. The cell proliferation was detected by Edu staining after 24 h (Scalebar, 100 μm) E. The cell migration was detected by wound healing assay after 24 h (Scalebar, 100 μm) F. The cell invasion was observed by Transwell assay after 24 h (Scalebar, 100 μm). Data were analyzed using the one-way analysis of variance followed by Tukey’s post hoc test; ^*p < 0.05, ^**p < 0.01, ^***p < 0.001