Cells were cultured and treated in 6-well dishes. Protein synthesis was assessed using the OPP protein synthesis kit (Invitrogen, C10457). Cells were incubated with 10 μM OPP in culture medium at 37°C for 1 hour, followed by fixation in 0.5 mL 10% neutral-buffered formalin (Sigma-Aldrich, HT501128) at 4°C for 15 minutes, permeabilized in 0.1% saponin (Sigma-Aldrich, 47036) in 500 μL PBS (Sigma-Aldrich, P4417) at room temperature for 5 minutes, and washed once with PBS. Fixed cells were resuspended and incubated in 500 μL freshly prepared OPP reaction cocktail (440 μL reaction buffer, 10 μL copper protectant, 1.25 μL picolyl azide, 50 μL reaction buffer additive) in the dark at room temperature for 30 minutes, washed with 0.5 mL rinse buffer, resuspended in PBS, and kept on ice before FACS analysis.

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