FAMK-1 catalytic activity and presence in the late secretory pathway contributes to fertility, embryogenesis, and posthatching development. (A) Phylogenetic tree of Fam20 family kinases. (B) Immunoblot of WT (N2) extract and extract prepared from famk-1Δ worms. α-Tubulin serves as a loading control. (C) Fertility and embryonic lethality analysis of individual worms of the indicated genotypes. (D) Phenotypes of posthatching famk-1Δ larvae. See also Fig. S1 B. (E) Immunoblot of WT and D387A mutant FAMK-1::mCherry expressed from single-copy transgene insertions. The blot was probed using an anti-FAMK-1 antibody. Actin serves as a loading control. (F) Fertility and embryonic lethality analysis for the indicated conditions. (G) Top: Schematics of WT and KDEL mCh::FAMK-1 (with mCherry inserted after signal peptide [SP]). Bottom: Immunoblot of extracts prepared from the indicated strains probed using an anti-FAMK-1 antibody. Actin serves as a loading control. (H) Fertility and embryonic lethality analysis for the indicated conditions. Error bars are the SD; *****, P < 0.0001 (t test).