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. 2014 Jul 1;127(13):2920–2933. doi: 10.1242/jcs.148130

Fig. 2.

Fig. 2.

Knockdown of p27 promoted cell migration. (A–C) GFP–p27 was ectopically expressed in p27−/− MEFs by using an adenovirus delivery method (A). At 24 h post-infection, the wound-healing assay (B) and transwell assay (C) were conducted to confirm the role of p27 in regulation of cell migration. Data show the mean±s.d. (three independent experiments); *P<0.05 (between Ad-GFP-p27- and Ad-GFP-infected cells). (D) Two sets of shRNA targeting different sequences of mouse p27 mRNA were stably transfected into p27+/+ MEFs, and the knockdown efficiency was determined by western blotting. Densitometric quantification of p27 expression is shown. (E,F) The wound-healing assay (E) and transwell assay (F) were used to determine the cell migration capability of shRNA-p27 and non-silencing control transfectants. (G,H) Cells were pretreated with Mitomycin C (10 µg/ml) for 3 h, and the wound-healing assay (G) and transwell assay (H) were conducted to detect the effect of p27-specific shRNA on cell migration. (I) Two sets of p27-specific shRNA were stably transfected into mouse epidermal Cl41 cells, and the knockdown efficiency was determined by western blotting. Densitometric quantification of p27 expression is shown. (J,K) The wound-healing assay (J) and transwell assay (K) were used to determine the cell migration capability of shRNA-p27 transfectants and non-silencing control transfectants. Data show the mean±s.d. (three independent experiments); *P<0.05 (between shRNA p27 and non-silencing control transfectants).