Analysis of intracellular trafficking activity of Leishmania cells expressing Myo21ΔUBA1-GFP and Myo21ΔUBA2-GFP.
Endocytic internalization of FM4-64 in (A) Myo21ΔUBA1-GFP expressing cells and (B) Myo21ΔUBA2-GFP expressing cells. Cells were incubated with FM4-64FX for 10 min before washing and suspending in fresh medium. Thereafter, aliquots of cells were taken at 0 min, 30 min, 60 min and 120 min time point. Adhered and fixed cells were stained with DAPI (blue) to visualize nucleus (N) and kinetoplast (K); FM4-64 dye is in red. Scale bar—2 μm. (C). Quantitative analyses of Myo21ΔUBA1-GFP and Myo21ΔUBA2-GFP expressing cells showing percent of total cells which trafficked FM4-64 dye beyond the nucleus in 60 min (n = 43 and 36 for Myo21ΔUBA1-GFP and Myo21ΔUBA2-GFP expressing cells, respectively, from three independent experiments), compared to Myo21-GFP expressing cells.