Fig. 2

Che-1 is highly phosphorylated. A A multiple sequence alignment of five mammalian Che-1 proteins. Below each position of the protein sequence alignment is present a key denoting conserved sites (*), sites with conservative replacements (:), sites with semi-conservative replacements (.) and sites with non-conservative replacements (). B Total cell extracts from HeLa cells treated or not with λ-PP (see Methods) were separated onto parallel 2D-Gel electrophoresis. C Schematic representation of Che-1 point mutations (see Methods) at the indicated sites. D 2D-Gel electrophoresis (top) and representative WB (bottom) of HeLa cells transiently transfected with Myc-Che-1 or Myc-Che-1 3S. E Representative immunofluorescence images (left) and WB analysis (right) of HCT116 cells transfected with the indicated expression vectors. Scale bar, 10 μm. F WB analysis with the indicated antibodies of total, cytoplasmic and nuclear fractions from HCT116 cells transfected as in D