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. 2015 Oct 22;10(10):e0139888. doi: 10.1371/journal.pone.0139888

Fig 5. ManNAz trafficking is regulated by PLD2.

Fig 5

(A) Cells were pulse-labeled for 1 h with ManNAz, washed and chased for 1 or 3 h. Cells were permeabilized to investigate ManNAz localization inside the cells. After 1 h of chase, ManNAz was localized juxtanuclearly in all cell lines. However, only PLD2CA and RBL-2H3 cells had cytoplasmatic vesicles containing ManNAz associated with the Golgi region (arrow). After 3 h of chase, all cell lines had ManNAz labeling in the Golgi complex as well as in cytoplasmatic vesicles in the same region (arrow). (Bars: 10μm). (B) In PLD2CA cells ManNAz concentration inside the cells was higher when compared to the other cells during the time periods analyzed. P<0.0001 for 1 and 3 hours. (C) Cells were pulse-labeled for 1 h with ManNAz, washed and chased for 24 h. Cells were permeabilized to investigate ManNAz localization inside the cells. ManNAz was localized juxtanuclearly and inside cytoplasmatic vesicles (arrows). (Bars: 10μm). (D) When the amount of ManNAz inside the cells was measured, there was more ManNAz labeling after 24 and 36 h of chase in PLD2CI cells than in RBL-2H3 and PLD2CA cells. P = 0.0463 for 24 hours. P = 0.0214 for 36 hours. For confocal microscopy, ManNAz was coupled to Alexa 488.