ZFIN ID: ZDB-FIG-200314-54
Rojas-Benítez et al., 2020 - Elongator Subunit 3 (Elp3) Is Required for Zebrafish Trunk Development. International Journal of Molecular Sciences   21(3) Full text @ Int. J. Mol. Sci.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage Range: 4-cell to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Long-pec

Figure 1

Elp3 morphants present a ventrally-curved tail. (A) Schematic representation of the anti-codon loop of a tRNA, indicating position 34 (red arrow) where 5-methoxycarbonylmethyl-2-thiouridine (mcm5s2U) is located. Anti-codon is depicted in green. (B) In situ hybridization of Elp3 mRNA in 4 cell (upper) and 27 hpf (lower) embryos. Scale bar 0.7 mm. (C) Saccharomyces cerevisiae mutants for ELP3 (ΔELP3) were transformed with an expression plasmid that contained zebrafish Elp3 (zElp3). BY4741 was used as control. Image representative of three independent experiments. (D) Western blot detecting Elp3 in 48 hpf samples from control and morphant embryos (MO-Elp3); β-actin was used as loading control. Samples from embryos injected with translation- (ATG) and splicing-blocking (SP). (E) Liquid chromatography-mass spectrometry (LC-MS) analysis of tRNA modification. Peak area values for mcm5s2U are shown normalized to the of purified tRNA for each sample (n = 3, * p < 0.05). (F) qPCR for UPR targets BiP, Chop and Atf4. Fold change in mRNA levels comparing morphant embryos to controls; t-test was performed for statistical analysis. *** p < 0.005). (G) Representative images of control, morphants and MO + ELP3 mRNA injected embryos by 48 hpf (images representative of at least six independent experiments, in which at least 100 embryos were analyzed).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
elp3 WT control Long-pec whole organism WB
WT standard conditions 4-cell whole organism ISH
Prim-5 whole organism ISH
WT + MO1-elp3 control Long-pec whole organism WB
WT + MO2-elp3 control Long-pec whole organism WB
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-elp3 control Long-pec whole organism elp3 expression decreased amount, abnormal
WT + MO1-elp3 standard conditions Long-pec post-vent region curved ventral, abnormal
Long-pec trunk curved ventral, abnormal
WT + MO2-elp3 control Long-pec whole organism elp3 expression decreased amount, abnormal
WT + MO2-elp3 standard conditions Long-pec post-vent region curved ventral, abnormal
Long-pec trunk curved ventral, abnormal
Acknowledgments:
ZFIN wishes to thank the journal International Journal of Molecular Sciences for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Int. J. Mol. Sci.