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Kinetics of MAL-FLAG internalization. COS-7 cells transfected with MAL-FLAG were incubated for 1 h at 4°C with anti-FLAG mAb and then fixed (a) or incubated at 37°C for the indicated times to allow MAL-FLAG internalization (b–g). Cells were permeabilized (b–g) or not (a) with 0.2% Triton X-100 and incubated with fluorescent secondary antibodies. (g and h) Double-label immunofluorescence analysis of MAL-FLAG internalized for 50 min (g) and steady-state TGN38 (h). Bar, 10 μm.
