|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 8. An active form of ILK (S343D) blocks the inhibitory effect of EphA1. (A) HEK293 (293) WT-EphA1–GFP and 
SAM-EphA1–GFP cells were transfected with wild-type ILK-myc (myc), starved of serum for 12 hours and then stimulated with ephrin-A1–Fc or control Fc (1 µg/ml) for 15 minutes. Cells were fixed and immunostained by anti-myc antibody (red). DAPI is blue; GFP is green. Note the colocalization of EphA1-GFP and ILK-myc in the merged images even before ephrin-A1–Fc stimulation in WT-EphA1–GFP cells (see the area with an arrowhead in the inset). Scale bar: 20 µm. (B) The same set of cells as is shown in A, with additional transfection with ILK-S343D, was subjected to spreading assays. The number of spreading cells over that of myc-positive cells was calculated. Data show means ± s.d. of three independent experiments. (C) ILK and RhoA activities in HEK293 WT-EphA1–FLAG cells transfected with wild-type ILK-myc (WT) or ILK-S343D–myc (S343D) were measured by immunoblot analysis as described in the Materials and Methods. (D) HEK293 WT-EphA1–GFP cells transfected with vector alone (lanes 1, 2) or ILK-S343D–myc (lane 3) were subjected to cell migration assays as described in Fig. 6E in the absence (lane 1) or presence (lanes 2, 3) of ephrin-A1–Fc.
|
