Phalloidin distribution

In order to check if the toxin was able to exhibit activity toward cytoskeleton of neuroblastoma cells, Louzao et al. stained filamentous actin (F-actin) of the cells with Oregon Green 514 Phalloidin and detected the fluorescence with LSC and confocal microscopy. Neuroblastoma cells incubated with 30 jM gambierol for 4 h, did not show modifications in the amount of F-actin or its distribution, indicating a cellular integrity in treated cells (Figure 28.16). Same results were observed with CTX3. 

To reveal the host cells, a fluorescent marker such as phalloidin can be added at this step. Phalloidin is a toxin from the toadstool Death Cap Amanita phalloides) that binds F-actin and is used to reveal the distribution of actin filaments in permeabilized cells. 

Figure 9.3 Distribution of focal adhesion proteins on RGD nanopattemed surfaces. Embryonic rat fibroblasts were seeded onto nanopattemed surfaces with either a 58- or 108-nm separation distance between RGD ligands. The localization of focal adhesion proteins vincuhn and paxillin were visualized using immunofluorescence at 3 and 24 h post-seeding. Cells seeded on the 58-nm nanopattemed surfaces showed significantly higher degrees of cell spreading compared with those on the 108-nm surfaces. Actin filaments in the fibroblasts were stained using an immunofluorescent phalloidin stain [64],...

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