JCS01057 Supplemental Data

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• Supplemental Figures -

  Fig. S1. G3DEGF expression reduced the number of cells arrested in G1 phase after serum withdrawal. (A) Cells were cultured in DMEM containing 10% FBS for 24 hours at 20% cell density, and little difference in cell cycle progression was observed between G3DEGF- and vector-transfected U87 cells. (B) After serum withdrawal from the cultures, the G3DEGF-transfected cells exhibited delayed G1 phase arrest in the cell cycle compared with the vector-transfected cells. The experiments were repeated three times.

  Fig. S2. Expression of G3DEGF altered cell morphology in serum-free culture. Cells were seeded in DMEM without serum and maintained for 24 hours, 48 hours and 8 weeks. At 24 hours, vector-transfected U87 cells had much longer processes, whereas G3DEGF-transfected U87 cells were less elongated. At 48 hours, vector-transfected cells continued to extend processes but most of the G3DEGF-transfected cells had detached from the plates and died. After 8 weeks of culture, vector-transfected cells had migrated, connected to each other and survived, but no G3DEGF-transfected cells survived. The experiment was repeated five times.

  Fig. S3. Effect of EGF on cell rounding. Cells were cultured overnight in DMEM containing 10% FBS. The medium was then replaced with DMEM alone or DMEM containing 10 ng ml^–1 EGF. After 5 days, cell morphology was examined and photographed. EGF enhanced cell rounding in vector-transfected U87 cells. The experiments were performed in three wells and repeated three times.