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| Figure 2: Experimental design of a cell-based high-throughput assay for identifying PCSk9 inhibitors. HepG2 cells were seeded in 2×105 /ml in 96-wells plate. Vehicle (PBS) or D374Y-PCSK9 protein (13.4 nM) was added in the culture medium with pooling 5 compounds into each well (200 nM final conc.) from the NINDS library. The positive pools in duplication were broken down to each individual compound. Each individual positive compound was then subject to the secondary screen assay by western blot. |
