Figure 3: TbMTase37 is a nucleolar protein.

(a) Western blot analysis was performed on total, nuclear (N) and cytoplasmic (C) protein fractions using polyclonal rabbit antibodies against TbMTase37, Nog1 (a nuclear/nucleolar marker) and Enolase (cytoplasmic marker). Anti-Enolase antibody (α-Enolase) was used as a control for cytoplasmic contamination in the nuclear fractions and anti-Nog1 antibody (α-Nog1) was used as a control for nuclear contamination of cytoplasmic fraction. Blots were generated with equal volumes of the same lysate on different gels and cropped panels represent the signal observed at the expected size of each protein. (b) Immunofluorescent localization using antibodies against NOG1 (α-NOG1) and anti-His antibody (α-His) to detect the His-tagged TbMTase37. DAPI (blue) was used to detect the nuclear (N) and mitochondrial kDNA (K). Fluorescent-labeled secondary antibodies were used to detect TbMTase37 (red) and NOG1 (green). DIC represents light microscope view of a single cell. Merged images are superimpositions of each single combination or all three channels together performed with ImageJ (NIH). Yellow fluorescence is indicative of co-localization of NOG1 and TbMTase37.