Fig. 5
Overexpression of CRABP2 promotes EMT, metastasis and invasion of ER− breast cancer by inactivating Hippo pathway. a Analysis of the YAP, p-YAPs127, Lats1, p-Lats1T1079 and Mst2 expression in BT549 and MDA-MB-231 cells stably expressing CRABP2 (Flag-CRABP2) and vector (Con) by Western blotting. b The express of YAP was analyzed in BT549 and MDA-MB-231 cells stably expressing CRABP2 (Flag-CRABP2) and vector (Con) by subcellular fractionation. Western blotting of GAPDH and LaminA/C were used as controls for cytoplasmic and nuclear fractional purity, respectively. c TEAD luciferase activity was analyzed in BT549 and MDA-MB-231 cells stably expressing CRABP2 (Flag-CRABP2) and vector (Con) were examined . d Analysis of CRABP2, Lats1, E-cadherin, ZO-1 and Vimentin expression in BT549 and MDA-MB-231 cells stably expressing CRABP2 (Flag-CRABP2) and vector (Con), with or without Lats1 (Flag-Lats1) by Western blotting. (e) Wound-healing, (f) migration and (g) invasion experiments were conducted in BT549 and MDA-MB-231 cells stably expressing CRABP2 (Flag-CRABP2) and vector (Con), with or without Lats1 (Flag-Lats1). The percent of wound closure and migratory and invasive cells numbers were counted. P-values were calculated by the Student’s t-test. Data are the mean ± S.D. of three independent experiments performed in triplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001