Figure 1

Imaging of IFP in Leishmania cells and Western blot analysis. A) Fluorescence images of Leishmania cells expressing IFP-His or ANAC42-His fusion protein were taken with a Cy5.5 filter set (665 ± 22.5 nm excitation and 725 ± 25 nm emission) using deconvolution microscopy. Illumination with white and infrared light simultaneously visualizes Leishmania cells and infrared signal, whereas infrared light detects infrared signal only. Infrared fluorescence signal is only detected in IFP-expressing cells, but not in cells expressing IFP-free negative control protein ANAC42-His. B) Several Leishmania cell lines (twelve IFP-His and eight ANAC42-His clones) were randomly selected and molecular weights of the expressed fusion proteins - 37 kDa for IFP-His and 33 kDa for ANAC42-His - were determined by SDS-PAGE analysis. Pre-stained markers were labeled on the membrane with a blue ball pen resulting in green-fluorescing dots upon excitation at 800 nm using the Odyssey Infrared Imaging System.