Table 1

Zebrafish tremblor ^tc318 heterozygotes were bred in the Tg(myl7:EGFP) background and raised as previously described (Westerfield, 2000 ). Embryos were raised at 28.5°C and staged as previously described (Kimmel et al., 1995 (link)). For Cn or proteasome inhibition, embryos were treated with 10 μM FK506 (Sigma-Aldrich, St. Louis, MO) or 50 μM MG132 (Sigma-Aldrich) at 24 hpf. The morpholino-modified antisense oligonucleotides targeting the translation initiation sites of murf1a and murf1b (Table 1, Gene Tools) were microinjected at the 1- to 2 cell stage (8 ng each). This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols of the University of California, Los Angeles. The protocol was approved by the Chancellor's Animal Research Committee of the University of California, Los Angeles.

Glγ-giardin expression was decreased using a 25-mer morpholino for the Glγ-giardin open reading frame (ORF) from the start codon (Table 1; Gene Tools Llc, Philomath, OR, USA). The control morpholino, a non-specific oligomer, was used as a control (Table 1). Morpholinos were added to 5 × 10⁶ cells at a final concentration of 100 μM by electroporation. The transfected cells were grown for 48 h, and then analyzed for level of Glγ-giardin by western blot and IFA as described above.