Figure 4
Figure 4. p27Kip1 depletion, Skp2 overexpression, and Cdh1 knock down abolished cell-adhesion–mediated p27Kip1 induction and cell-cycle arrest. (A) The p27Kip1 expression in control siRNA entering RISC-transfected (siCont.RISC[ + ]) or siP27Kip1 RNA-transfected (siP27) Jeko-1 cells in suspension (Sus) versus 12 hours of HS-5 adhesion (HS5-Ad) analyzed by Western blot, and (B) corresponding cell-cycle distribution in suspension (Sus) versus HS-5 adhesion (HS5-Ad) analyzed by flow cytometry. (C) The Skp2 and p27Kip1 expression in nontransfected [pCont(−)] Jeko-1 cells in suspension (Sus) versus HS-5 adhesion (HS5-Ad) or stably mock (pCont( + ))- or plasmid Skp2-transfected (pSkp2) Jeko-1 cells in suspension versus to HS-5 adhesion for 12 hours analyzed by Western blot and (D) corresponding cell-cycle distribution in stably mock- or plasmid Skp2-transfected Jeko-1 cells in suspension (pCont( + )-Sus or pSkp2-Sus) versus HS-5 adhesion (pCont( + )-Ad or pSkp2-Ad) for 12 hours analyzed by flow cytometry. (E) Cdh1, Skp2, and p27Kip1 expressions in nonsilencing siControl RNA-transfected (siCont) or siCdh1 RNA-transfected (siCdh1)-Jeko-1 cells in suspension (Sus) versus HS-5 adhesion (HS5-Ad) for 12 hours analyzed by Western blot, and (F) corresponding cell-cycle distribution in control siRNA- or Cdh1 siRNA-transfected Jeko-1 cells in suspension (siCont-Sus or siCdh1-Sus) versus HS-5 adhesion (siCont-Ad or siCdh1-Ad) for 12 hours analyzed by flow cytometry. Each blot is representative of 3 experiments, and the bar graphs show means plus or minus SD of 3 experiments.

p27Kip1 depletion, Skp2 overexpression, and Cdh1 knock down abolished cell-adhesion–mediated p27Kip1 induction and cell-cycle arrest. (A) The p27Kip1 expression in control siRNA entering RISC-transfected (siCont.RISC[ + ]) or siP27Kip1 RNA-transfected (siP27) Jeko-1 cells in suspension (Sus) versus 12 hours of HS-5 adhesion (HS5-Ad) analyzed by Western blot, and (B) corresponding cell-cycle distribution in suspension (Sus) versus HS-5 adhesion (HS5-Ad) analyzed by flow cytometry. (C) The Skp2 and p27Kip1 expression in nontransfected [pCont(−)] Jeko-1 cells in suspension (Sus) versus HS-5 adhesion (HS5-Ad) or stably mock (pCont( + ))- or plasmid Skp2-transfected (pSkp2) Jeko-1 cells in suspension versus to HS-5 adhesion for 12 hours analyzed by Western blot and (D) corresponding cell-cycle distribution in stably mock- or plasmid Skp2-transfected Jeko-1 cells in suspension (pCont( + )-Sus or pSkp2-Sus) versus HS-5 adhesion (pCont( + )-Ad or pSkp2-Ad) for 12 hours analyzed by flow cytometry. (E) Cdh1, Skp2, and p27Kip1 expressions in nonsilencing siControl RNA-transfected (siCont) or siCdh1 RNA-transfected (siCdh1)-Jeko-1 cells in suspension (Sus) versus HS-5 adhesion (HS5-Ad) for 12 hours analyzed by Western blot, and (F) corresponding cell-cycle distribution in control siRNA- or Cdh1 siRNA-transfected Jeko-1 cells in suspension (siCont-Sus or siCdh1-Sus) versus HS-5 adhesion (siCont-Ad or siCdh1-Ad) for 12 hours analyzed by flow cytometry. Each blot is representative of 3 experiments, and the bar graphs show means plus or minus SD of 3 experiments.

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