Supplemental Material

This article contains the following supporting material:

• Figure 1 - Detection of endogenous SMAP2 protein by immunoblot analysis.
  The lysates from the untransfected HeLa and Cos-7 cells were processed for immunoblot analysis. The anti-SMAP2 antibody was raised in a rabbit, using the peptide sequence (aa 1-15) as an antigen. The endogenous SMAP2 was detected as a 47kDa band. The extreme left lane represents the SMAP2-transfected HeLa cells. Unfortunately, this antibody did not work in immunofluorescent detection of endogenous SMAP2 protein.

• Figure 2 - A possibility that SMAP2 forms a dimmer.
  HeLa cells were transfected by SMAP2 (aa 1-428) and the cell lysate was processed for immunoblot analysis. The anti-SMAP2 antibody could detect not only 47kDa but also 94kDa bands. These two bands represent the specific immunoreaction, since the inclusion of excess amount of antigen peptide into the antibody solution could abolish these two bands. Thus, the 94kDa band likely represents a homodimer of SMAP2, although a possible complex between SMAP2 and an unknown molecule of 47kDa cannot be excluded. Another observation supporting a dimer formation of SMAP2 came from yeast two hybrid screening. When a cDNA library was screened using SMAP2 as a bait, a cDNA encoding a portion of SMAP2 was obtained as a positive clone.