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Fig. 6 | Breast Cancer Research

Fig. 6

From: Rack1 mediates tyrosine phosphorylation of Anxa2 by Src and promotes invasion and metastasis in drug-resistant breast cancer cells

Fig. 6

Increased expression of Anxa2WT or Anxa2Y23D in Rack1-silenced cells recovered cell invasion ability. a, b Increased expression of Anxa2WT, Anxa2Y23A, and Anxa2Y23D in Rack1 stably silenced MCF-7/ADR cells (a) and MDA-MB-468/EPR cells (b). Lentivirus expressing Anxa2WT, as well as its two mutants, were used to infect Rack1 stably silenced cells, in which the expression of Rack1 has been stably transfected with an shRNA targeting its noncoding region. Then, the cells were lysed and analyzed by Western blotting using anti-Anxa2, anti-GFP, and anti-Rack1 antibodies. β-actin was used as a loading control. c, d Transwell assays showed that overexpression of Anxa2WT or Anxa2Y23D, not Anxa2Y23A, in Rack1-silenced MCF-7/ADR (c) and MDA-MB-468/EPR cells (d) partially rescued the cell migration and invasion abilities. Data are shown as mean ± SD; n = 6. Statistical analysis was performed by one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and nsP > 0.05 indicate no statistical significance. e Src inhibitor KX2-391 efficiently inhibited the expression of pY23-Anxa2 in Anxa2WT-expressing cells, while had no significant effect on the level of pY23-Anxa2 in Anxa2Y23D-expressing cells as detected by Western blotting. f Transwell assay showed that the migration ability in Anxa2WT-expressing cells can be quenched by Src inhibitor, while Src inhibition has little effect on the migration ability in Anxa2Y23D-expressing cells. Data are shown as mean ± SD; n = 6. Statistical analysis was performed by one-way ANOVA. *P < 0.05, **P < 0.01, ****P < 0.0001, and nsP > 0.05 indicate no statistical significance

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