Fig 4. Knockdown of Src-1 or Twist1 expression in CNE-1 suppressed colony formation, anchorage-independent growth, cell migration and invasion.
(A) 500 CNE-1 cells transduced with sh-Src-1, sh-Twist1 or sh-NC control were seeded in 6-well-plates, allowed grow for 2 weeks and stained with crystal violet. (B) Colony numbers from A. (C) 5000 CNE-1 cells transduced with sh-Src-1, sh-Twist1 or sh-NC control were seeded in soft agar and cultured for 3 weeks, then stained with 1 mg/ml MTT. (D) colony numbers from C. (E) 1×105 CNE-1 cells transduced with sh-Src-1, sh-Twist1 or sh-NC control were used for transwell cell invasion assay, then stained with Giemsa dye and imaged after incubating for 24hr. (F) percentage of invasion cells from E comparing with sh-NC. (G) CNE-1 cells transduced with sh-Src-1, sh-Twist1 or sh-NC control were used for scratch wound healing assay, photos of the plates were took at 0h and 48h. Scale bars = 50um.(H) Relative cell migration from G analyzed by Digimizer software system. (I) cell proliferation of sh-Src-1, sh-Twist1 or sh-NC transduced CNE-1 cells was evaluated by cell viability assay. ***P<0.05 comparing with sh-NC group.