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Cells were maintained in an incubator at 37 C and 5% CO 2 incubator and seeded onto 6-well cell culture plates before transfection

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As a complement to removing EPN-1, we tested the signaling activities of forms of both LAG-1 and APX-1 that were mutated or truncated to block their ability to interact with Epsin per se or with all cytosolic proteins respectively (Figure 1C)

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