Cellular localization of GFP-tagged ISCL.
(A–E) Log phase WT [pXG-GFP-ISCL] parasites were stained with MitoTracker Red580 and subjected to fluorescence microscopy. (A) DIC; (B) Hoechst staining; (C) MitoTracker staining; (D) GFP fluorescence; (E) Merge of C and D. (F) GFP-ISCL is functional. Whole cell lysates from iscl− or iscl− transfected with pXG-GFP-ISCL were tested for neutral SMase activity as described and a representative TLC image is shown. Positive control (+): 0.1 unit of B. cereus SMase; negative control (−): boiled WT lysate.