|
|
Blood, Vol. 107, Issue 11, 4514-4523, June 1, 2006
Both carboxy-terminus NES motif and mutated tryptophan(s) are crucial for aberrant nuclear export of nucleophosmin leukemic mutants in NPMc+ AML
Blood Falini et al.
107: 4514
Supplemental materials for: Falini et al
Files in this Data Supplement:
- Figure S1 (JPG, 26.5 KB)
-
NIH 3T3 cells were transiently transfected with either pEGFP-C1 vector or pEGFP-C1-NPMwt or pEGFP-C1-NPMmA, and, 24 hours later, lysed in ice-cold NP-40 lysis buffer as indicated in Methods section. Proteins were separated by SDS-PAGE, electroblotted on PVDF membrane and subjected to Western Blot analysis using anti-GFP mouse monoclonal antibody (Roche, Indianapolis, IN) (upper panel). The same membrane was stripped and re-blotted with either the anti-NPM mouse monoclonal antibody, Cl. 376 (middle panel) or the rabbit polyclonal antibody, Sil-C, raised against the mutated form of NPM protein (lower panel). To note, while the anti-NPM, Cl. 376 antibody was able to recognize either the wild-type or the mutated form of the GFP-NPM fusion protein (64 kDa MW), the Sil C antibody was able to recognize only the mutated protein. These data confirm the specificity of the Sil-C antibody for NPM mutant protein in an overexpression system.
| |
