FIGURE 7.
p27kip1 repression induced by HIPK2 depletion reduces cell motility in p53-null cells. H1299-CV and H1299i cells were transfected with the control pcDNA3.1 vector or with vectors encoding wild-type p27-HA, p27S10A-HA, or p27S10D-HA. A, TCEs from transfected cells were analyzed by WB for the indicated proteins. B, cells from the same transfection analyzed in A were seeded on 5-mm pore size Transwell filters and allowed to migrate toward 10% FCS. After 48 h, cells on the underside of the filters were fixed, stained, and analyzed as indicated under “Experimental Procedures.” Upper panel, one representative experiment of five independent ones is shown. Cell motility was visualized at 40× magnification. Lower panel, migrated cells were quantified and expressed as mean ± S.D. (lines) of five independent experiments, assuming the value of control vector-transfected H1299 cells was equal to 1. Statistical significance was assessed for the differences between H1299-CV cells transfected with control or p27S10A-HA vectors (*) and between H1299i cells transfected or not with p27-HA (**) and p27S10D-HA (***) (p < 0.05).