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. 2016 Mar 1;6:22. doi: 10.3389/fcimb.2016.00022

Figure 5.

Figure 5

AmVs interact with the ER in mammalian host cells. (A,B) The AmV interacts with the ER and ER-derived vesicles are delivered into its lumen. A. marginale infected RF/6A cells were screened with antibodies against calreticulin (A), PDI (A), or derlin-1 (B). The regions that are demarcated by hatched lined boxes indicate the regions magnified in the insets that are demarcated by solid lined boxes. (C–E) Derlin-1-positive vesicles are present within the AmV in close association with A. marginale bacteria. A. marginale infected RF/6A cells were screened with antibodies against derlin-1 and Msp5 and visualized using LSCM. (D) Z-stack series shows derlin-1-labeled vesicles within the AmV. Successive focal planes of the region in (C) that is demarcated by a hatched line box are presented. (E) 3D rendering of the Z-stack series presented in (D) shows derlin-1-positive vesicles in close proximity to intravacuolar A. marginale organisms. Host cell nuclei and bacteria were stained blue with DAPI. Scale bars, 5 μm. Results shown are representative of two experiments with similar results.