Cells were plated in Lab-Tek II Chamber slides (Labclinics) 24 hours after siRNA transfection. SNAP labeling was initiated 48 hours after transfection (24 hours after plating). For quench-chase-pulse experiments, cells were incubated with 5 μM SNAP-Cell Block (S9106S, New England Biolabs) for 30 min at 37°C. After two PBS washes, cells were incubated in medium for 30 min, followed by two washes. Cells were incubated in medium for the chase period (6 to 7 hours). Cells were incubated with 1 μM TMR-Star (S9105S, New England Biolabs) for 30 min, washed twice, incubated in medium for 30 min, washed twice, preextracted for 5 min in 0.2% Triton X-100/PBS on ice, and fixed for 10 min in 4% PFA.

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