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Cryo-EM sample preparation and data acquisition The purified wild-type hTauT protein was concentrated to about A280=20 (11 mg ml) to prepare the apo samples and incubated at 4 C for 2 h with 2 mM taurine, 5 mM -alanine, or 2 mM P4S separately to prepare substrate-bound samples and inhibitor-bound samples