Fig. 4. Accumulation of neddylated proteins in DEN1^null mutants. (A-C) Transgenes UAS-myc-Nedd8GG-CFP (A), UAS-myc-Nedd8AA-CFP (B) and UAS-myc-mNedd8 (C) are expressed by the GAL4 driver ms1096 in wing pouches (outlined). Nedd8 expression, as shown by Myc staining (purple), was enhanced in GFP-negative DEN1^null clones (arrows) for Myc-Nedd8GG-CFP (A) and Myc-mNedd8 (C), but not for Myc-Nedd8AA-CFP (B). Right panels show merged images. Mutant clones were located by the absence of the nuclear GFP signal (in left panels, see Materials and Methods for genotypes), which could be differentiated from the CFP signal (for the fusion proteins) by their emission spectra using Carl Zeiss LSM Meta 510 confocal microscope.