Localization of VSGΔgpi in bloodstream cells. Bloodstream cells expressing either 117Δgpi (A to H) or 221Δgpi (I to P) were incubated for 2 h in the absence (A, B, E, F, I, J, M, and N) or presence (C, D, G, H, K, L, O, and P) of FMK024 (117Δgpi, 2 μM; 221Δgpi, 20 μM). Fixed and permeabilized cells were stained as follows: panels E, G, M, and O, anti-BiP (red) and anti-VSGΔgpi (green); panels F, H, N, and P, anti-p67 (red) and anti-VSGΔgpi (green). All samples were counterstained with DAPI to reveal the nucleus (n) and the kinetoplast (k). Three channel merged images are presented in which colocalization is represented as yellow. The corresponding differential interference contrast-DAPI merged images are presented above each panel. The insets in panels F to H and panels N to P are the corresponding single channel images (p67, red; VSGΔgpi, green) in the region of the lysosome.