Fig. 3. Disorganization of HA cross-linking in normal fibroblasts promoted breast cancer malignancy.

A Primary tumor cells from MMTV-PyMT mice were stained with cell tracker (green) and then co-cultured with or without corresponding normal fibroblasts (NFb^Control and NFb^TSG6−) in the tumor–stromal assay (TSA). As shown in the left image, tumor cells (green) were in the middle of TSA and surrounded by fibroblasts (red, illustrated by staining vimentin). After 72 h, the proliferation of tumor cells was determined by staining Ki67. The top panel shows the representative images of tumor cells (green) with nuclei (DAPI). The down panel shows the Ki67 expression (red) in corresponding nuclei. B After stimulation with 50% conditioned media (CM) of NFb^Control or NFb^TSG6−, the apoptosis of tumor cells was evaluated by the TdT-DAB apoptosis assay. Red arrows indicate the apoptotic tumor cells. C The colony formation abilities of tumor cells treated with NFb^Control CM or NFb^TSG6− CM. D The statistical histograms of Ki67 percentage, apoptotic percentage, and colony count of tumor cells in different groups. E The migration and invasion abilities of tumor cells. All experiments were repeated three times with similar results. Error bars represent mean ± SD values. Statistical analysis was performed using Student’s t test. **p < 0.01, ***p < 0.001 (vs NFb^Control or NFb^Control CM), ^##p < 0.01 (vs Control).