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. 2014 May 22;6(5):1644–1666. doi: 10.3390/toxins6051644

Figure 4.

Figure 4

Single endo/lysosome analysis. The endo/lysosomal escape of Alexasaporin in the presence of SA1641 was monitored in the case of single endo/lysosomes. (A) ECV-304 cells were incubated with Alexasaporin at 1000 nM and 37 °C for 3 h, washed, and then, 10 µg/mL SA1641 were added to cells as indicated in Figure 3. AlexaSaporin is visualized in green fluorescence. A total of 16 single endo/lysosomes was selected and constantly monitored from t = 2050 s to t = 3290 s. For each endo/lysosome, a region of interest (numbered 1–16 in the fluorescence picture) comprised of the surroundings of the endo/lysosomes, but excluding the vesicle itself, was defined; (B) ECV-304 cells were continuously monitored for 7200 s after the addition of 10 µg/mL of SA1641. Single endo/lysosome analysis is presented as a relation of time (from t = 2400 s to t = 4200 s) vs. the fluorescence intensity of the released Alexasaporin. Each curve represents the fluorescence intensity increase in each region of interest, corresponding to the amount of Alexasaporin that escapes from each single endo/lysosome.