Supplemental Figures

This article contains the following supporting material:

• Figure S1 - Endogenous TRAF7 and c-Myb are localized in both cytoplasm and nucleus in DND39 B cells. DND39 cells were permeabilized with Triton X-100, immunostained with antibodies against the protein indicated above the panel, and visualized by FITC- and rhodamine-conjugated secondary antibodies, using confocal microscopy. DNA was stained with TOTO3. c-Myb, TRAF7, and DNA are shown in green, red, and blue, respectively. In the right most panels, the signals for both proteins and DNA are superimposed. Typical staining pattern of single cell is shown below with higher magnification.
• Figure S2 - Over-expressed TRAF7 in CV-1 cells sequesters c-Myb in the cytoplasm. (A) Localization of over-expressed 2KR mutant of c-Myb in the nucleus. CV-1 cells were transfected with the wild-type c-Myb expression plasmid and the β-galactosidase expression plasmid. Transfected cells were permeabilized with digitonin (upper panels) or Triton X-100 (lower panels), immunostained with antibodies against the proteins shown above, visualized by FITC- and rhodamine-conjugated secondary antibodies, and analyzed by confocal microscopy. DNA was stained with TOTO3. β-Galactosidase, c-Myb (2KR), and DNA are shown in red, green, and blue, respectively.In the right most panels, the signals for both proteins and DNA are superimposed. (B) (C) TRAF7 sequesters wild-type c-Myb, but not the 2KR mutant, in the cytoplasm. CV-1 cells were transfected with the wild-type (B) or 2KR mutant (C) c-Myb expression plasmid and the TRAF7 expression plasmid, and analyzed as described in Figure 7C.
• Figure S3 - TRAF7 sequesters SUMO1 and c-Myb in the cytoplasm. CV-1 cells were transfected with plasmids expressing wild-type c-Myb (A, B) or the 2KR mutant (C), SUMO1, and TRAF7, permeabilized, immunostained, and analyzed as described in Figure 8B-D.