Figure 3
TGFαL3SEA D227A inhibits tumour growth in vitro. (A) A431 cells were seeded into a 96-well plate at a density of 2.5 × 104 cells/well. Following an overnight incubation period, various ratios of effector cells and target cells were co-cultured with A431 cells in the presence of 5 ng/μl rSEA(squire) or TGFαL3SEAD227A(black diamond). Viable tumour cells were determined using an MTT assay, and the data were given as a percentage of tumour cell growth inhibition (TCGI). (B) A431 cells were plated in a 96-well plate at a density of 2.5 × 104 cells/well, followed by treatment with the indicated concentrations of TGFαL3SEAD227A (black diamond) and rSEA (square) at an E/T ratio of 30:1 in vitro. Viable tumour cells were determined as described above (A). (C) This figure indicates the alignment of the third loop of human TGFα and mice TGFα. (D)The protein level of EGFR in A431, B16 and HEK293T cells was detected by western blotting. (E) B16 cells, plated in a 96-well plate at a density of 2.5 × 104, were treated identically to the A431 cells with the indicated dose of TGFαL3SEAD227A (black diamond), rSEA(square), and BSA (triangle).