For adult protein lysates, equal numbers of adults (50–60) from each condition were picked into microcentrifuge tubes containing 33 μL Laemmli sample buffer. Samples were then boiled for 10 min, centrifuged for 10 min at 14000 x RPM at RT, and the supernatants were transferred to fresh tubes. Equal volumes were loaded per lane.

For embryo lysates, ~ 100–200 adults from each condition were collected, and fresh bleach solution (20% sodium hypochlorite + 0.25 M NaOH in ultrapure H2O) was added to each tube to dissolve the adults. After ~ 8 min with gentle mixing (or until adults were dissolved), embryos were washed once with M9, resuspended in 15 μL M9, and then the number of eggs was counted using a dissecting scope. Samples were then mixed with 2x Laemmli sample buffer, boiled, and centrifuged before loading. Equal numbers of eggs (~ 800) were loaded per lane.

Lysates were loaded into a freshly prepared 8% SDS-PAGE. Gels were stained using Coomassie brilliant blue and imaged using a ChemiDoc™ XRS System (Bio-Rad Laboratories Inc., Hercules, CA).

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