Fig. 2

PTPN20 enhances the metastatic ability of TNBC cells. (A) Western blotting demonstrated knockdown of PTPN20 expression following incubation with shRNA targeting PTPN20-3’UTR or PTPN20 plasmids in the MDA-MB-231 and SUM159PT cell lines. GAPDH was used as a loading control. (B) Representative micrographs and the percentage of wound closure of the indicated cells by wound healing assay. (C) Representative micrographs and quantification of the invasiveness of the indicated cells in the transwell matrix penetration assays. (D) Western blotting demonstrated overexpression of PTPN20 following incubation with PTPN20 plasmids in the BT549 and MDA-MB-468 cell lines. GAPDH was used as a loading control. (E) Representative micrographs and the percentage of wound closure of indicated cells with overexpression of PTPN20 by wound healing assay. (F) Representative micrographs and quantification of the invasiveness of indicated cells with overexpression of PTPN20 in the transwell matrix penetration assays. (G) Representative micrographs of tumors in the indicated groups. n = 6 for each group. (H) Tumor volumes were measured on the indicated days in the indicated groups. n = 6 for each group. (I) Average weight of excised tumors from the indicated mice. n = 6 for each group. (J) Lung metastasis burden of xenografted animals was monitored weekly using bioluminescent imaging (BLI). Shown are BLI images of representative mice on day 40 after injection. The color scale depicts the photon flux (photons per second) emitted from the metastasis cells. (K) Representative images and H&E staining of lung tissues. Black irregular circles marking the metastatic nodules. Scale bar, 50 μm. (L) The metastasis index was calculated (the ratio of lung metastatic nodules number/tumor weight) in the indicated mice. n = 6 for each group. In (B), (C), (E) and (F), error bars represent the mean ± SD of three independent experiments. In (B) and (C), Kruskal-Wallis test was used. In (E), (F), (I) and (L), Mann-Whitney U test was used. In (H), One-way repeated-measures ANOVA test was used. ***P < 0.001, **P < 0.01, *P < 0.05