Figure 3: Schematic of the presented auxin-mediated depletion protocols.

After bleaching adult worms to isolate embryos, synchronized hatched L1s can be placed on regular NGM food plates until they reach the desired stage. For Basic Protocol 1, L4-stage or adult worms (approximately on Day 4-5, depending on growth temperature) can be placed on auxin-NGM plates to initiate depletion; depletion in this method occurs on the scale of hours. For faster protein depletion, worms that are fed on regular NGM plates until adulthood can be soaked in auxin-containing media for up to an hour (Basic Protocol 2) or dissected into auxin-containing media to analyze isolated embryos (Basic Protocol 3) before proceeding with experiments.