Whole-mount and Cryosectioned Immunofluorescence Protocols

Whole-mount IF experiments were completed as previously described (Gerlach and Wingert, 2014 (link); Kroeger et al., 2017 (link); Marra et al., 2017 (link), 2019c (link); Chambers et al., 2020a (link),b (link)). For cilia and basal bodies, anti-tubulin acetylated diluted 1:400 (Sigma-Aldrich, T6793) and anti γ-tubulin diluted 1:400 (Sigma-Aldrich, T5192) were used, respectively. Cryosectioned samples were completed as previously described (Gerlach and Wingert, 2014 (link)). For cilia and basal bodies, anti-tubulin acetylated diluted 1:1000 (Sigma-Aldrich, T6793) and anti γ-tubulin diluted 1:400 (Sigma-Aldrich, T5192). For cell polarity, animals were fixed in Dent's solution, and anti-aPKC diluted 1:500 (Santa Cruz Biotechnology, 2300359) was used to mark apical surface and anti-Na⁺K⁺ ATPase diluted 1:35 (Developmental Studies Hybridoma Bank, 528092) for a basolateral marker. See Table S1 for antibody details.