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To elucidate the potential mode of action of MAP-based cytolytic fusion proteins in living cells, colocalization assayswere performed using EGFR+ A549 cells transfected with a SNAP-tagged tubulin DNA construct. Expressed SNAP-tagged tubulin molecules were labeled with SNAP-Cell TMR-Star (green). EGF-MAPf151 was used as a representative for MAP-based CFPs. MAPf151 was detected using mouse–anti-human Tau and goat–anti-mouse Alexa Fluor 647 antibodies (red). DAPI was used to counterstain the nucleus (blue). Using confocal fluorescence microscopy and a tubulin polymerization assay, it could be shown that MAP colocalized with and stabilized microtubules. For more details, see article by Hristodorov and colleagues on page 2194.