Figure 2.
PIPKIγ and IQGAP1 cooperate to regulate migration. (A) MDA-MB-231 cells were transfected with the indicated siRNA for 48 h. Knockdown was confirmed by immunoblotting with the indicated antibodies (top). Using Transwell, 10% serum-induced migration (middle) and invasion through 2 mg/ml Matrigel (bottom) were measured. (B) Cells were transfected with the indicated DNA and siRNA combinations for 24 h. Expression level was analysed by immunoblotting with the indicated antibodies (top). Migration and invasion were measured as in (A) (bottom). (C) PIPKIγi1 was expressed in HeLa tet-off cells by removing doxycycline from media for 24 h. Protein expression and cell motility were measured as above. Data are shown as mean±s.d. for four independent experiments. (D) Cells maintained in suspension were either plated on 10 ng/ml collagen I or kept in suspension for 30 min. Serum-starved cells were treated with or without 10% serum for 15 min. Endogenous IQGAP1 was IP’ed and associated PIPKIγ was analysed by immunoblotting. COL, type I collagen; IQ1, IQGAP1; S, serum; Sus, suspension. (E) Iqgap1 KO MEFs were stably reconstituted with the indicated IQGAP1 proteins, and four different modes of migration were measured with a Transwell (top right). Protein expression was analysed by immunoblotting (top left). Conditions used for treating Transwells (bottom). Data are shown as mean±s.d. of four independent experiments. Data above are representative of at least four independent experiments.
Source data for this figure is available on the online supplementary information page.
