Fig. 5.

Re-expression of Rack1^WT, not Rack1^Y246F, rescued Anxa2 phosphorylation and invasion ability in drug-resistant cancer cells. a, b Rescued expression of Rack1^WT, not the Src binding-deficient Rack1^Y246F mutant, recovered Anxa2 phosphorylation in MCF-7/ADR cells (a) and in MDA-MB-468/EPR cells (b). Rack1 expression was stably silenced by using shRNA targeting its noncoding region. Then, the cells were infected with lentivirus expressing flag-tagged Rack1^WT or Rack1^Y246F and selected by using hygromycin B. The cell lysates were analyzed by Western blotting with anti-Rack1, Flag, Anxa2, and phospho-Anxa2 antibodies. c, d Re-expression of Rack1^WT, not Rack1^Y246F, recovered cell migration and invasion abilities. Transwell assay was used to analyze the migration and invasion abilities in MCF-7/ADR cells (c) and in MDA-MB-468/EPR cells (d). Representative images were captured at 200×. Statistical analysis was performed by one-way ANOVA. Statistical results were exhibited in the following panel. ****P < 0.0001 and ^nsP > 0.05 indicate no statistical significance