FIGURE 4.

Cytotoxic effect of batimastat is brought about by inhibition of sAPPα generation. A, CD18 cells were transfected with siRNA to APP or ADAM10 and Western blot analysis was performed using 6E10 and ADAM10 antibodies to confirm knockdown of the respective genes. B, APP or ADAM10 were knocked down in MiaPaCa2 cells and Western blot was used to confirm knockdown. The effect of ADAM10 and APP knockdown in CD18 (C) and MiaPaCa2 (D) cells was compared with batimastat treatment in a cytotoxicity assay using MTT reagent at A₅₄₀. CD18 (E) and MiaPaCa2 (F) cells were treated with 100 nm recombinant sAPPα alone or in combination with 5 μm batimastat for 24 h and changes in cell death was analyzed with the MTT reagent at A₅₄₀. Each experiment was performed three times, and (*)(#)(**) indicates significant difference with p value less than 0.05 from an unpaired two-tailed Student's t test.