Photoactivation and click chemistry were performed using a previously described method with some modifications (12, 39). Briefly, cell lysates containing the packaged pseudotyped viruses were incubated with 50 μM Y0, Y11, or Y12 or with DMSO for 30 min at room temperature. The lysates were then incubated with 5 μM Y18 for an additional 30 min and exposed to UV light (365 nm) for 10 min on ice. Azido-immobilized agarose and 10 mM l-ascorbic acid were then added sequentially. The azido-immobilized agarose was prepared by incubating excess biotin-azide (Invitrogen) with streptavidin (SA)–immobilized agarose (Invitrogen). The cycloaddition reaction (click chemistry) was initiated by adding 1 mM CuSO4, and the samples were incubated overnight at 4°C. The agarose was washed, and the captured target proteins were analyzed by Western blotting.

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