Figure 6. SKR-5 Localizes to the ER and Is Required for the Clearance of Misfolded Proteins in
ire-1;
daf-2 Mutants(A) Representative western blot of DAF-28::GFP and tubulin of day 2
cup-4 mutants and
ire-1;
daf-2 double mutants expressing a DAF-28::GFP transgene treated with
sel-11 RNAi or control RNAi. Both strains are coelomocyte-defective. Bar graph presents the tubulin-normalized levels of DAF-28::GFP ± SEM in three independent biological experiments. Asterisks mark Student's t test values of p < 0.01 compared to control RNAi treatment.(B) DAF-28::GFP levels were compared between
ire-1;
daf-2 double mutants and
ire-1;
daf-2;
skr-5 triple mutants on day 2 of adulthood. Both strains are coelomocyte-defective. DAF-28::GFP and tubulin levels were assessed by western blot (left) and by fluorescence. Bar graph presents the normalized mean levels of DAF-28::GFP ± SEM as assessed by fluorescence or by western blot measurement. The experiment averages more than three independent biological repeats. '' n '' indicates number of animals analyzed.(C) Eggs of the indicated genotypes were treated with 5 mg/ml tunicamycin. Percentage of eggs that developed into mature adults or L4 within 4 days ± SEM is shown. Each experiment was repeated independently with similar effects.(D)
skr-5 mutants treated with control RNAi and wild-type animals treated with
sel-11 RNAi developed from eggs into adults within 3 days.
skr-5 mutants treatedwith
sel-11 RNAi did not complete their development into adults even after 6 days.