Figure 5.

Ub^G76V-GFP turnover in adults is mediated by the UFD complex and the EGF/Ras/MAPK-signalling pathway. (A, B) Co-expression of Ub^G76V-GFP (green, top panels) and mRFP (red, bottom panels) proteins in C. elegans at either (A) 24 h or (B) 48 h after L4 stage for the indicated genotypes. (C) Relative levels of Ub^G76V-GFP and mRFP fluorescence are quantified as a ratio (AU indicates arbitrary units) at either 24 h (solid bars) or 48 h (stippled bars) after L4 stage for the indicated genotypes. Hypomorphic mutants for let-60, mek-2, and eor-2 show a modest change in the ratio; it is unclear whether this change is due to residual gene activity in the mutants or indicates that these factors do not exclusively mediate this effect. ^***P<0.0001, ^**P<0.001, ^*P<0.01 by two-way ANOVA/Bonferoni's post hoc comparisons between the 24 and 48-h time points for each genotype. For the statistical effect of genotype on the time-dependent outcome, P<0.0001 for all mutants compared with wild type. Error bars indicate s.e.m., N=15–25 animals per trial. (D) Ub^G76V-GFP and actin were detected by western blot at either 24 or 48 h after L4 for the indicated genotypes. Equivalent amounts of protein were loaded into each lane. Bar, 100 μm.