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The pulse-chase procedure for the chloroalkane penetration assay (CAPA). (i) The Halo-GFP-Mito (red) is cytosolically oriented and bind covalently to the ct-molecules (green). (ii) After washing, ct-dye (gray) chases unoccupied HaloTag and provides fluorescence upon binding. (iii) The fluorescent signal is quantified by flow cytometry and normalized fluorescence is plotted as a function of ct-molecule concentration.
