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. 2020 May 25;10:212. doi: 10.3389/fcimb.2020.00212

Figure 4.

Figure 4

Shiga toxin 2 can be taken up by Gb3-negative cells within microvesicles. (A) CHO-native cells were incubated with CFSE-stained microvesicles (green dye) containing Stx2:Alexa555 (Stx2-pos MVs, red dye) and stained with DAPI. Fluorescence microscopy showed that microvesicles were associated with CHO-native cells and that some of the microvesicles were positive for Stx2:Alexa555. (B) DLD-1 cells were incubated with microvesicles positive for Stx2:Alexa488 (Stx2-pos MVs, green dye). 3D-SIM imaging showed that fluorescence corresponding with Stx2:Alexa488 was present within DLD-1 cells (as determined by z-stack acquisition) when delivered via microvesicles. (C) CHO-native cells were incubated with free Stx2:Alexa555 and stained with DAPI. No fluorescence was seen in CHO-native cells that had been given Stx2:Alexa555 in free form. (D) DLD-1 cells were incubated with free Stx2:Alexa488. No fluorescence was seen in DLD-1 cells that had been given Stx2:Alexa488 in free form. Two independent experiments were performed and representative results are presented.