Fig. 5

B591 abolishes tumor growth in vivo by eliminating CSCs. a MDA-MB-231 xenografts were treated with vehicle or B591 for 2 weeks, then the residual tumor tissue was harvested and dissociated into single cells for subsequent CSCs assays according with the flow chart. b–d Tumor weight and mice body weight were measured every 2 days during treatment. In the end of treatment, tumor tissues were collected and weighted (n = 8). e The CSCs markers of tumor cells were analyzed by flow cytometry. Only the live and non-mouse cell population was gated and analyzed. Data from three mice were plotted. f Mammosphere formation assay was carried out with the single cells. The relative mammosphere-formation ability is shown as percentage, and data represent mean ± SD. g The single cells from vehicle and 16 mg/kg of B591 group were replanted into NOD/SCID mice according to the indicating number, and 3 months later, tumors were counted for calculating the tumor-initiating frequency. h The CSCs markers and the downstream substrates of PI3K pathway were analyzed using the lysates from the primary xenografts by western blot analysis. ^**p < 0.01, difference versus vehicle-treated control group