ZFIN ID: ZDB-FIG-191230-1467
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:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: 1-cell to Shield
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: 1-cell to Shield

Fig 7 Up-regulation of the MARCKS family members and its interaction protein Hsp70.3 over-compensates the genetic loss of <italic>marcksb</italic>.

(A, B) RT-qPCR analysis on the expression of marcksa, marcksb, marcksl1a, marcksl1b and hsp70.3 in the embryos indicated in the Fig at 1-cell stage (A) and shield stage (B). The data were presented as scatter plots with bar representing median value relative to respective transcript levels measured in wildtype embryos. “*”: P < 0.01, from Student’s t-test. (C) Overexpression of marcksa, marcksl1a, marcksl1b or hsp70.3 partially rescued the dorsalization defects in marcksb morphants. “n” represents the number of embryos we observed. (D) Co-immunoprecipitation revealed that Hsp70.3 had high binding affinity with Marcksb and moderate binding affinity with Marcksl11a and Marcksl1b, but relative low binding affinity with Marcksa. TCL: total cell lysis. The molecular mass of Marcks-myc is around 70KD. (E) The expression of szl was remarkably reduced in MZmarcksb injected with either hsp70_MO or marcksa_l1a_l1b_MO. The embryos are at shield stage and lateral view with dorsal to the right. (F) The percentage of embryos with normal-like and severely decreased expression of szl. “n” represents the number of embryos we observed. (G) Hsp70 interacts with Marcksa, Marcksl1a and Marcksl1b to maintain sufficient level of extracellular Bmp2b in MZmarcksb. (a-c) Compared with MZmarcksb embryos (a), knockdown of either hsp70 (b) or a combination of marcksa, marcksl1a and marcksl1b (c) remarkably reduced the level of extracellular Bmp2b in MZmarcksb. (H) Quantitative measurement of secreted Bmp2b in embryos of MZmarcksb, MZmarcksb injected with either hsp70_MO or marcksa_l1a_l1b_MO. The data were presented as scatter plots with median; “*”: P < 0.01, from Student’s t-test.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
hsp70.3 AB + MO1-marcksb standard conditions Shield whole organism RTPCR
marcksbihb199/ihb199 standard conditions 1-cell whole organism RTPCR
Shield whole organism RTPCR
marcksbihb199 standard conditions 1-cell whole organism RTPCR
Shield whole organism RTPCR
marcksb AB + MO1-marcksb standard conditions Shield whole organism RTPCR
marcksbihb199/ihb199 standard conditions 1-cell whole organism RTPCR
Shield whole organism RTPCR
marcksbihb199 standard conditions 1-cell whole organism RTPCR
Shield whole organism RTPCR
marcksl1a AB + MO1-marcksb standard conditions Shield whole organism RTPCR
marcksbihb199/ihb199 standard conditions 1-cell whole organism RTPCR
Shield whole organism RTPCR
marcksbihb199 standard conditions 1-cell whole organism RTPCR
Shield whole organism RTPCR
marcksl1b AB + MO1-marcksb standard conditions Shield whole organism RTPCR
marcksbihb199/ihb199 standard conditions 1-cell whole organism RTPCR
Shield whole organism RTPCR
marcksbihb199 standard conditions 1-cell whole organism RTPCR
Shield whole organism RTPCR
szl marcksbihb199/ihb199 standard conditions Shield whole organism ventral region ISH
marcksbihb199/ihb199 + MO1-hsp70.3 standard conditions Shield whole organism ventral region ISH
marcksbihb199/ihb199 + MO1-marcksa + MO1-marcksl1a + MO2-marcksl1b standard conditions Shield whole organism ventral region ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-marcksb standard conditions Shield whole organism hsp70.3 expression decreased amount, abnormal
Shield whole organism marcksb expression decreased amount, abnormal
marcksbihb199/ihb199 standard conditions 1-cell whole organism marcksb expression decreased amount, abnormal
1-cell whole organism marcksl1a expression increased amount, abnormal
1-cell whole organism marcksl1b expression increased amount, abnormal
Shield whole organism hsp70.3 expression increased amount, abnormal
Shield whole organism marcksb expression decreased amount, abnormal
marcksbihb199/ihb199 + MO1-hsp70.3 standard conditions Shield whole organism ventral region szl expression decreased amount, abnormal
Shield whole organism ventral region szl expression decreased distribution, abnormal
marcksbihb199/ihb199 + MO1-marcksa + MO1-marcksl1a + MO2-marcksl1b standard conditions Shield whole organism ventral region szl expression decreased amount, abnormal
Shield whole organism ventral region szl expression decreased distribution, abnormal
marcksbihb199 standard conditions 1-cell whole organism marcksb expression decreased amount, abnormal
1-cell whole organism marcksl1a expression increased amount, abnormal
Shield whole organism hsp70.3 expression increased amount, abnormal
Shield whole organism marcksb expression decreased amount, abnormal
Shield whole organism marcksl1a expression increased amount, 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.