ZFIN ID: ZDB-FIG-191230-1466
Ye et al., 2019 - Marcksb plays a key role in the secretory pathway of zebrafish Bmp2b. PLoS Genetics   15:e1008306 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage: Shield
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Shield to Prim-5

Fig 6 MZ<italic>marcksb</italic> showed genetic compensation on BMP signaling.

(A, B) Injection of marcksb_MO in MZmarcksb did not cause defect of dorsalization. (A) morphological observation of MZmarcksb and marcksb_MO injected MZmarcksb. Up panels, field view of embryos at 12 hpf; lower panels, representative embryos at 30 hpf. (B) WISH analysis showed that the transcriptional levels of downstream targets of BMP signaling—szl and ved were at comparable level between MZmarcksb and marcksb_MO injected MZmarcksb. (C) The overall intensity of p-Smad1/5/9 at ventral region in MZmarcksb was slightly increased when compared with wildtype. (D) The normalized fluorescent intensity of P-Smad1/5/9. Error bars of light blue and light red show S.E.M. (E) The extracellular mCherry-Bmp2b was slightly increased in the MZmarcksb compared with wildtype embryos. (F) Quantitative measurement of secreted Bmp2b in wildtype and MZmarcksb embryos. The data were presented as scatter plots with median; *”: P < 0.01, from Student’s t-test. (G) Although knockdown of chd in wildtype embryos only resulted in moderate ventralization (V1-V2), knockdown of chd in MZmarcksb embryos resulted in severe ventralization (V3-V4). V1-V4, ventralization type 1 to type 4 embryos; the statistical data are shown in the bar graphs with the number of observed embryos indicated right. (H) After injection of chd_MO, the up-regulation of BMP signaling activity was much more robust in MZmarcksb than that in wildtype embryos shown by WISH analysis of szl (a vs b) and ved (c vs d). The embryos are at shield stage and animal-pole view with dorsal to the right; (I) The percentage of embryos with normal-like, increased, and dramatically-increased expression of szl and ved. “n” represents the number of embryos we observed.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
szl AB standard conditions Shield whole organism ventral region ISH
AB + MO1-chrd standard conditions Shield whole organism ventral region ISH
marcksbihb199/ihb199 standard conditions Shield whole organism ventral region ISH
marcksbihb199/ihb199 + MO1-chrd standard conditions Shield whole organism ventral region ISH
marcksbihb199/ihb199 + MO1-marcksb standard conditions Shield whole organism ventral region ISH
ved AB standard conditions Shield whole organism ventral region ISH
AB + MO1-chrd standard conditions Shield whole organism ventral region ISH
marcksbihb199/ihb199 standard conditions Shield whole organism ventral region ISH
marcksbihb199/ihb199 + MO1-chrd standard conditions Shield whole organism ventral region ISH
marcksbihb199/ihb199 + MO1-marcksb standard conditions Shield whole organism ventral region ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab13-smad IHC AB standard conditions Shield whole organism ventral region
IHC marcksbihb199/ihb199 standard conditions Shield whole organism ventral region
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-chrd standard conditions Shield whole organism ventral region szl expression increased distribution, abnormal
Shield whole organism ventral region ved expression increased distribution, abnormal
Prim-5 whole organism ventralized, abnormal
marcksbihb199/ihb199 standard conditions Shield whole organism ventral region Ab13-smad labeling increased amount, abnormal
Shield whole organism ventral region szl expression increased distribution, abnormal
Shield whole organism ventral region ved expression increased distribution, abnormal
marcksbihb199/ihb199 + MO1-chrd standard conditions Shield whole organism ventral region ved expression increased distribution, abnormal
Shield whole organism ventral region szl expression increased distribution, abnormal
Prim-5 whole organism ventralized, exacerbated
marcksbihb199/ihb199 + MO1-marcksb standard conditions Shield whole organism ventral region ved expression increased distribution, abnormal
Shield whole organism ventral region szl expression increased distribution, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.