Preparation of crude P. falciparum nuclei for proteomic analysis. (a) Visual assessment of the crude nuclear preparation. Intact parasites (control) and isolated nuclei were stained with DAPI and analyzed by DIC and fluorescence microscopy. Scale bar 2 μm. (b) TEM analysis of isolated nuclei shows intact as well as damaged nuclei and very little contamination from other organelles. Arrows indicate individual nuclei. (c) Immunoelectron microscopy verifies the identity of nuclei. Anti-H3K4me3 antibodies were used to label nuclei in intact cells (left panel) and in the nuclear preparation (right panel) (arrows). (d) 1D-SDS PAGE analysis of all protein fractions obtained from trophozoites, visualized by silver staining. (e) Analysis of trophozoite cytoplasmic and nuclear fractions by western blot.
Oehring et al. Genome Biology 2012 13:R108 doi:10.1186/gb-2012-13-11-r108