Fig. 4

SHH002-hu1 inhibits Bevacizumab-induced transwell, migration and EMT of TNBC cells via abating Wnt/β-catenin pathway. a. Microscopic views from transwell assay to estimate MDA-MB-231/MDA-MB-468 cells invasion following 24 h treatment of Bevacizumab/Bevacizumab + SHH002-hu1/Bevacizumab + FH535, bar = 100 μm. b. Quantitative analysis of (a) by Image J. c. Photomicrographs of cell migration from wound healing assay in MDA-MB-231/MDA-MB-468 cells treated with Bevacizumab/Bevacizumab + SHH002-hu1/Bevacizumab + FH535, bar = 100 μm. d. Quantitative analysis of (c) by Image J. The quantitative analyses of transwell invasion assay and wound healing assay were based on the mean of the 5 regions of each group. Data were presented as the mean ± SD, n = 5, ^*p < 0.05, ^**p < 0.01. e. SHH002-hu1 inhibited Bevacizumab-induced EMT in TNBC cells. MDA-MB-231/MDA-MB-468 cells were incubated with Bevacizumab/Bevacizumab + SHH002-hu1/Bevacizumab + FH535 for 24 h. Then the results of western blot analysis for EMT marker proteins were shown. f. TOP/FOP ratio in MDA-MB-231/MDA-MB-468 cells treated for 24 h with Bevacizumab/Bevacizumab + SHH002-hu1/Bevacizumab + FH535. SHH002-hu1/FH535 inhibited the β-catenin/TCF-4 transcriptional activity induced by Bevacizumab significantly. Data were presented as the mean ± SD, n = 3, ^*p < 0.05, ^**p < 0.01. g. SHH002-hu1 suppressed the activation of Wnt/β-catenin pathway induced by Bevacizumab. The results of western blot analysis for the expression of β-catenin and downstream oncoproteins were shown