Figure 5. Wild-type Sur8, but not phosphomimetic mutations, mediates FGF2-induced cellular transformation, migration, and invasion.

(A-F) GFP-Sur8-WT, GFP-Sur8-T71E, or GFP-Sur8-S297D which were rescued in DLD-1 Sur8-KO cells were treated with PBS (control) or FGF2 and assessed for transformation, proliferation, migration, and invasion potential, and were subjected to immunoblot and RT-PCR analyses. Foci-forming assays were conducted for 2 weeks. Quantifications of colony number are shown in (A). Normal (B) and anchorage-independent (C) cell growth rates were determined by MTT and soft agar assays, respectively. Cells were scratched and allowed for wound closure for 48 hours after treatment with or without FGF2. Representative images of wound closure and quantification of relative wound closure efficiency are shown in (D). Invasion assays were performed using matrigel-coated chambers, and the relative quantification of invaded cells were measured as shown (E). Colonies and cells were stained with crystal violet (A, C-E). Statistical values were ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. Scale bars, 500 μm. WCEs were subjected to immunoblot analysis and total RNAs were subjected to RT-PCR analysis (F).