Research Paper Volume 13, Issue 1 pp 1032—1050

SPOCK1/SIX1axis promotes breast cancer progression by activating AKT/mTOR signaling

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Figure 3.

SPOCK1 promotes cellular invasion, metastasis and the EMT in vitro and in vivo. (A) A scratch wound-healing assay was used to determine the effects of SPOCK1 on BC cell motility. (B) Results of a transwell migration assay (a) and a Matrigel invasion assay (b) for cellular invasion. The mean number of cells in five fields per membrane is shown (×200). (C) Representative images of gross and hematoxylin and eosin (H&E) staining and relative numbers of lung surface metastatic foci detected in each group (*P<0.5, **P<0.01). The scale bar is 100 μM and 50 μM. (D) Representative images showing the morphological changes in the indicated cell lines. (E) The heat maps of the correlation between SPOCK1 and EMT markers in the same cohort. (F) Positive relationships for SPOCK1 and EMT markers were showed on GEPIA2. (G) The expression of EMT markers was detected by immunofluorescence staining in BC cells. The scale bar is 20 μM. (H) The expression of epithelial markers (E-cadherin and ZO-1) and mesenchymal markers (Vimentin, Snail, Slug, Twist, S100A4 and ZEB1) was determined by western blot analysis. GAPDH was used as a loading control. (I) IHC staining for E-cadherin and Vimentin protein in tumor specimens from xenografts (200×). The P values were obtained using Mann-Whitney U tests or t-tests (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001). All results are from three independent experiments. The error bars represent the SD.