Antibody staining was performed as described previously (33). Antibodies were used for immunostaining after fixation with 2% paraformaldehyde (PFA) for 5 min and cold methanol for 3 min (for all except ARLE-14 and MET-2 antibodies, which was fixed with 2% PFA for 10 min and methanol for 3 min). For mutant configurations, an on-slide WT sample was included, marked with a single-copy ZEN-4::GFP tag. On-slide controls allowed better quantitation between different genotypes or stages. A description of how antibodies against endogenous MET-2 or ARLE-14 were generated can be found in a separate section.

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