Fig. 2

RBCK1 overexpression inhibited TNBC cell progression in vitro and in vivo. A RBCK1 over-expression efficiency by plasmids. We overexpressed RBCK1 in TNBC cells via lenti-virus infect system. The level of Flag-RBCK1 protein was detected by western blot. β-Actin was used as the internal reference. B–C Overexpression of RBCK1 inhibits the proliferation of TNBC cells. The metabolic activity of cells at the specified time points after transfection was determined by CCK-8 analysis. Each group was analyzed three times. *P < 0.05; **P < 0.01; ***P < 0.001 for cell growth comparisons. D–E Overexpression of RBCK1 inhibited cell migration in TNBC cells. MDA-MB-231 and BT549 cells with stable expression of RBCK1 or empty vector. Calculate the number of cells and the number data showed as ± SD. *P < 0.05; **P < 0.01; ***P < 0.001 for cell comparisons. F–G Overexpression of RBCK1 inhibited cell invasion in TNBC cells. MDA-MB-231 and BT549 cells with stable expression of RBCK1 or empty vector. Calculate the number of cells and the number data showed as ± SD. *P < 0.05; ** P < 0.01; ***P < 0.001 for cell comparisons. H–I Wound healing experiment of MDA-MB-231 and BT549 cells with stable expression of RBCK1 or vector. Wound closure was quantified at a specified point in time. *P < 0.05; **P < 0.01; ***P < 0.001 for cell comparisons. J–K Overexpression of RBCK1 increased apoptosis in TNBC cells. MDA-MB-231 and BT549 cells with stable expression of RBCK1 or empty vector were stained with PI and Annexin V. Each group was analyzed three times. *P < 0.05; **P < 0.01; ***P < 0.001 for comparisons. L–M Overexpression of RBCK1 inhibited the expression of CD24-CD44 + in TNBC cells. MDA-MB-231 and BT549 cells with stable expression of RBCK1 or empty vector were stained with anti-CD44-FITC and anti-CD24-PE. Each group was analyzed in three times. *P < 0.05; **P < 0.01; ***P < 0.001 for comparisons. N–P Overexpression of RBCK1 inhibited the tumor growth of MDA-MB-231 cells in a xenograft model. Tumor growth photographs, curves and weights are shown in panels N, O and P, respectively. The data are showed as ± SD. *P < 0.05; **P < 0.01; ***P < 0.001 for cell comparisons