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Notch processing was not affected by JNJ-40418677 treatment in a cell-free Notch assay. A recombinant substrate mN99-Flag was incubated with an enriched γ-secretase preparation (Winkler et al., 2009) from HeLa cells as described, and NICD levels were analysed by Western blotting utilizing an anti-Flag antibody. With up to 10 µM JNJ-40418677, no inhibition of Notch processing was observed, while NICD production was completely blocked by 1 µM DAPT.
