Comparing the effects of Ddx19B or Dicer1 knockdown by Trim-Away or morpholinos in zebrafish embryos. Zebrafish embryos were co-injected with TRIM21 and antibodies (anti-Ddx19B antibody, anti-Dicer1 antibody, or control IgG) or injected with morpholinos (ddx19b-MO, Dicer1-MO, or Con-MO) at 0 hpf. a The level of Ddx19B or Dicer1 was determined by western blotting at the indicated time points. b Lateral view of eye size at the indicated time points. c Lateral view of the body axis at 2 dpf. d Eye diameter in embryos injected with different doses of TRIM21/anti-Ddx19B antibody or TRIM21/IgG was measured at the indicated time points; zebrafish numbers are indicated in brackets. e The level of miR-26a and let-7a was determined at the indicated time points. f Lateral view of embryos at the indicated time points. The data in d and eare shown as the mean ± SD of three independent experiments, *P ≤ 0.05, **P ≤ 0.01, NS indicates P > 0.05

Degradation of Mitfa occurs 18 h after injection of TRIM21 and anti-Mitfa antibody. aEGFP expression plasmid or control pcDNA3.1 plasmid was injected into one-cell embryos. TRIM21 and anti-EGFP antibody were then co-injected into EGFP-expressing embryos or pcDNA3.1 plasmid-injected embryos at 24 hpf. The levels of EGFP, anti-EGFP antibody, and TRIM21 were analyzed by western blotting at different time points. bMitfa expression in zebrafish embryos was determined at different time points by western blotting. c One-cell embryos were co-injected with TRIM21 and an anti-Mitfa antibody or nonspecific IgG and analyzed for the levels of Mitfa, TRIM21, and IgG heavy chain at different time points by western blotting. d, e One-cell zebrafish embryos were injected with mitfa morpholinos (mitfa-MO) or control morpholinos (Con-MO) or co-injected with TRIM21/anti-Mitfa antibody or TRIM21/nonspecific IgG. Mitfa knockdown was confirmed at 22 hpf by western blotting; the arrow indicates IgG heavy chain (d). Lateral view of the embryos at different time points; embryos treated with N-phenylthiourea (PTU) were used as positive controls (e)