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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: CRABP2 regulates invasion and metastasis of breast cancer through hippo pathway dependent on ER status

Fig. 4

CRABP2 suppresses EMT, metastasis and invasion of ER+ breast cancer cells by activating Hippo pathway. a. Analysis of the YAP, p-YAPs127, Lats1, p-Lats1T1079 and Mst2 expression in T47D and MCF7 cells stably expressing CRABP2 shRNA (sh-CRABP2–1, sh-CRABP2–2) and shRNA (sh-NC) by Western blotting. b. The express of YAP was analyzed in T47D and MCF7 cells stably expressing CRABP2 shRNA (sh-CRABP1, sh-CRABP2–2) and shRNA (sh-NC) by subcellular fractionation. Western blotting of GAPDH and LaminA/C were used as controls for cytoplasmic and nuclear fractional purity, respectively. c The expression of YAP was analyzed in T47D and MCF7 cells expressing CRABP2 siRNA (si-CRABP2) and siRNA (si-NC) by immunofluorescence. Scalebar, 10 μm. d TEAD luciferase activity was analyzed in T47D and MCF7 cells stably expressing CRABP2 shRNA (sh-CRABP2–1, sh-CRABP2–2) and shRNA (sh-NC). e Analysis of CRABP2, Lats1, E-cadherin, ZO-1 and Vimentin expression in T47D and MCF7 cells stably expressing CRABP2 shRNA (sh-CRABP2–1, sh-CRABP2–2) and shRNA (sh-NC), with or without Lats1 (Flag-Lats1) by Western blotting. (f) Wound-healing, (g) migration and (h) invasion experiments were conducted in T47D and MCF7 cells stably expressing CRABP2 shRNA (sh-CRABP2–1, sh-CRABP2–2) and shRNA (sh-NC), 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

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