Fig. S8

(A-C) A knockdown of TTLL3 expression in zebrafish changes fish morphology and affects cilia in multiple organs. Morphology of control embryos (A) and TTLL3-ATG morphants (B-C) at 48 hpf that were released manually from the chorion after 24 hpf (B) or at 72 hpf released manually from the chorion after 48 hpf (C). (D) Results of an RT-PCR assay for TTLL3 mRNA using total RNA from control and morphants injected with 3 or 5 ng of TTLL3-SP MO. (E and F). Depletion of TTLL3 reduces the TAP952 signal of tubulin monoglycylation in pronephron cilia; a control embryo (E) and a TTLL3-ATG morphant (F). (G-N) TTLL3 depletion shortens cilia and reduces the TAP952-positive tubulin monoglycylation. G and H are images of neuromasts stained with 6-11 B-1 anti-acetylated α- tubulin antibodies in control (G) and a TTLL3-ATG morphant (H). (I-N) Double labeling of control embryos (I-K) and TTLL3-ATG morphants (L-N) with TAP952 (I and L) and polyE anti-polyglutamylation antibodies (J and M). (K and N) Merged images. Note a decrease in the level of tubulin monoglycylation and increase in the level of tubulin polyglutamylation in morphants.