Fig. 4
HA derived from HS5 stromal cells affected the growth of MDA-MB-231BO-GFP cells. a. The expression of three HA synthases, HAS1, HAS2, and HAS3, in HS5 cells was detected by qPCR. b. The interference efficiency of HAS2 siRNA was evaluated by qPCR in HS5 cells. c. The expression of HAS2 was detected by western blots after interference with HAS2 siRNA in HS5 cells. d. HA content in the supernatant was determined after HAS2 expression was downregulated in HS5 cells. (****p < 0.0001 compared with the NC siRNA group as measured by unpaired Student’s t test). e. The growth of HS5 cells after downregulation of HAS2 was detected by CCK-8 assays. F. A total of 3000 HS5-NC siRNA cells/well were plated in special 96-well plates. Twenty-four hours later, 0.375 × 103 MDA-MB-231BO-GFP cells were plated in the HS5 cell wells. MDA-MB-231BO-GFP cells were cultured alone as controls. After three days of culture, the fluorescence intensity of GFP was determined (****p < 0.0001 compared with the control group as measured by unpaired Student’s t test). G. After HAS2 in HS5 cells was knocked down by HAS2 siRNA, the cells were cocultured with MDA-MB-231BO-GFP cells for 72 h. The fluorescence intensity of GFP was measured. (**p < 0.01 compared with the HS5-NC siRNA group as measured by unpaired Student’s t test). Bars represent the mean ± SEM