ZFIN ID: ZDB-FIG-190716-4
Han et al., 2019 - Vitamin D Stimulates Cardiomyocyte Proliferation and Controls Organ Size and Regeneration in Zebrafish. Developmental Cell   48(6):853-863.e5 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Conditions:
Anatomical Term:
Stage Range: Day 4 to Adult
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: Day 4 to Adult

Fig. 4

Figure 4. ErbB2 Blockade Inhibits Vitamin D-Induced Cardiomyocyte Proliferation

(A) Selected functional cluster enrichment of differentially expressed genes. The activation score infers the activation score of the corresponding process.

(B) Selected common upstream regulators of differentially expressed genes in adult hearts after three Alfa injections. The p value measures the statistical significance of the overlap between differentially expressed genes and the genes under predicted control of a regulator, and the activation score infers the activation state and score of a regulator, either activating (positive) or inhibiting (negative).

(C) MIP images of dissected 4 dpf cmlc2:FUCCI hearts after 24-h treatment. Scale bar, 50 μm.

(D) Quantification of FUCCI+ CMs of 4 dpf cmlc2:FUCCI embryos after 24-h treatment. Mean ± SD, Mann-Whitney test and Student’s t test. ns, not significant; ∗∗∗p < 0.001. n = 11.

(E) MIP images of dissected 4 dpf hearts after 24-h treatment. Scale bar, 50 μm.

(F) Quantification of FUCCI+ CMs of 4 dpf embryos after 24-h treatment. Mean ± SD, Mann-Whitney test and Student’s t test. ∗∗p < 0.01; ∗∗∗∗p < 0.0001. n = 8–11.

(G) PCNA/Mef2 staining of ventricles from Alfa-injected zebrafishtreated with vehicle or AG1478 for 24 h, starting at 2 h after the last Alfa injection. Scale bar, 100 μm.

(H) Quantification of CM proliferation indices of experiments in (H). Mean ± SD, Student’s t test. n = 9.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
mCherry pd57Tg/+ ; pd302Tg/+ ; pd58Tg/+ control Day 4 cardiac muscle cell IFL
pd57Tg/+ ; pd302Tg/+ ; pd58Tg/+ chemical treatment by environment: tyrphostin AG 1478 Day 4 cardiac muscle cell IFL
pd57Tg/+; pd58Tg/+ chemical treatment by environment: alfacalcidol Day 4 cardiac muscle cell IFL
pd57Tg/+; pd58Tg/+ chemical treatment by environment: tyrphostin AG 1478, chemical treatment by environment: alfacalcidol Day 4 cardiac muscle cell IFL
Venus pd57Tg/+ ; pd302Tg/+ ; pd58Tg/+ chemical treatment by environment: tyrphostin AG 1478 Day 4 cardiac muscle cell IFL
pd57Tg/+; pd58Tg/+ chemical treatment by environment: tyrphostin AG 1478, chemical treatment by environment: alfacalcidol Day 4 cardiac muscle cell IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-mef2 IHC WT chemical treatment by injection: alfacalcidol Adult cardiac muscle cell
IHC WT chemical treatment by injection: tyrphostin AG 1478, chemical treatment by injection: alfacalcidol Adult cardiac muscle cell
Ab2-pcna IHC WT chemical treatment by injection: alfacalcidol Adult cardiac muscle cell
IHC WT chemical treatment by injection: tyrphostin AG 1478, chemical treatment by injection: alfacalcidol Adult cardiac muscle cell
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment by injection: alfacalcidol Adult cardiac muscle cell proliferation increased occurrence, abnormal
WT chemical treatment by injection: tyrphostin AG 1478, chemical treatment by injection: alfacalcidol Adult cardiac muscle cell proliferation occurrence, ameliorated
pd57Tg/+ ; pd302Tg/+ ; pd58Tg/+ control Day 4 cardiac muscle cell proliferation increased occurrence, abnormal
pd57Tg/+ ; pd302Tg/+ ; pd58Tg/+ chemical treatment by environment: tyrphostin AG 1478 Day 4 cardiac muscle cell proliferation occurrence, ameliorated
pd57Tg/+; pd58Tg/+ chemical treatment by environment: alfacalcidol Day 4 cardiac muscle cell proliferation increased occurrence, abnormal
pd57Tg/+; pd58Tg/+ chemical treatment by environment: tyrphostin AG 1478, chemical treatment by environment: alfacalcidol Day 4 cardiac muscle cell proliferation occurrence, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 48(6), Han, Y., Chen, A., Umansky, K.B., Oonk, K.A., Choi, W.Y., Dickson, A.L., Ou, J., Cigliola, V., Yifa, O., Cao, J., Tornini, V.A., Cox, B.D., Tzahor, E., Poss, K.D., Vitamin D Stimulates Cardiomyocyte Proliferation and Controls Organ Size and Regeneration in Zebrafish, 853-863.e5, Copyright (2019) with permission from Elsevier. Full text @ Dev. Cell