ZFIN ID: ZDB-FIG-160408-5
Batut et al., 2015 - Expression patterns of CREB binding protein (CREBBP) and its methylated species during zebrafish development. The International journal of developmental biology   59(4-6):229-34 Full text @ Int. J. Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: 14-19 somites
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: 14-19 somites

Fig. S2

Validation of CREBBP-methylated specific antibodies on zebrafish embryos. (A,B) Immunohistochemistry with CREBBPmethylated specific antibody at 18 ss using (A) R714me2a or (B) R742me2a antibody on embryos injected with Control morpholino (CTRMo) and CREBBPb morpholino (CREBBPbMo). Nuclei are stained in blue with To-Pro3 and a merged picture is shown. Expression of CREBBP-R714me2a and CREBBP-R742me2a is strongly reduced in CREBBPbMo- as compared to controlMo-injected embryos. Injections were performed at the 1-cell stage with 6 ng of CTRMo or CREBBPbMo (Gene-tools). Morpholino sequences were: CREBBPb 5′-GACTGTTGCCACCTGCCATGCCCAT-3′ and Control 5′-CCTCTTACCTCAGTTACAATTTATA-3′. (C) Knock-down of CREBBPb reduces CREBBPb protein level in 14 ss old zebrafish embryos. Embryos injected with either CTRMo or CREBBPbMo were collected at 14 ss and processed for western blot analysis to detect CREBBP expression. Tubulin was used as a loading control and a protein ladder is shown. Whole cell extracts from 20 embryos were classically prepared in 40 ml Laemmli sample buffer. 5 ml of each sample (4 embryos) were loaded per lane and subjected to SDS-PAGE analysis. The following antibodies were used: anti-CREBBP (1/200, A22, santa Cruz), anti a-Tubulin (1:5000, Sigma, T9026), anti-Rabbit IgG, HRP conjugate (1:50000, Promega, W4011) and anti-Mouse IgG, HRP conjugate (1:10000, Promega, W4021). Protein Ladder, PageRuler prestained Protein Ladder (Fermentas, SM0671).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
crebbpa Ab2-crebbpa WT control 14-19 somites somite IHC
Ab3-crebbpa 14-19 somites somite IHC
Ab3-crebbpa 14-19 somites whole organism WB
Ab2-crebbpa 14-19 somites whole organism WB
Ab2-crebbpa WT + MO3-crebbpa standard conditions 14-19 somites somite IHC
Ab3-crebbpa 14-19 somites somite IHC
Ab3-crebbpa 14-19 somites whole organism WB
Ab2-crebbpa 14-19 somites whole organism WB
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO3-crebbpa standard conditions 14-19 somites somite crebbpa expression decreased distribution, abnormal
14-19 somites somite crebbpa expression decreased distribution, abnormal
14-19 somites whole organism crebbpa expression decreased amount, abnormal
14-19 somites whole organism crebbpa expression decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal The International journal of developmental biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Int. J. Dev. Biol.