ZFIN ID: ZDB-FIG-161018-24
Olsen et al., 2016 - G9a and ZNF644 Physically Associate to Suppress Progenitor Gene Expression during Neurogenesis. Stem Cell Reports   7(3):454-70 Full text @ Stem Cell Reports
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Long-pec to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 5

znf644a and znf644b Morphant Retinas Exhibit Distinct Cellular Defects

(A) Immunostaining of retinal cross-sections of Vsx2 (red), Pax6 (red), Isl2b (green), or H3K9me2 (red) in WT or znf644a morphant embryos at indicated time points. Vsx2 expression at 48 hpf marks proliferating RPCs in the CMZ (gray arrows) and in a subpopulation of bipolar neurons in the central retina (yellow arrows). Pax6 expression marks amacrine cells and a subset of RGCs, and Isl2b expression marks a subset of RGCs. Differentiated retinal neurons with H3K9me2+ nuclei are highlighted.

(B and C) Immunostaining monitoring cleaved Caspase3 (red) in retinal cross-sections from WT, g9a, or znf644b morphant embryos at 48 or 72 hpf (B). Yellow arrows highlight Caspase3+ apoptotic cells. (C) Quantitation of Caspase3+ cells in WT, znf644a morphant, or znf644b morphant retinas with or without co-injection of p53-MO at 72 hpf. Quantitation represented as mean ± SD (n=3 for each group). *p < 0.05, **p < 0.005, ***p < 0.0005, Student′s t test.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
vsx2 Ab1-vsx2 AB standard conditions Long-pec ciliary marginal zone cell IHC
Ab1-vsx2 Long-pec retinal bipolar neuron IHC
Ab1-vsx2 AB + MO1-znf644a standard conditions Long-pec ciliary marginal zone cell IHC
Ab1-vsx2 Long-pec retinal bipolar neuron IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Qualifier Anatomy
Ab1-pax6 IHC AB standard conditions Long-pec amacrine cell
IHC Long-pec retinal ganglion cell
IHC Protruding-mouth retinal bipolar neuron
IHC AB + MO1-znf644a standard conditions Long-pec Not Detected amacrine cell
IHC Long-pec Not Detected retinal ganglion cell
IHC Protruding-mouth Not Detected retinal bipolar neuron
Ab5-casp3 IHC AB + MO1-ehmt2 standard conditions Long-pec retina
IHC Protruding-mouth retina
IHC AB + MO1-znf644b standard conditions Protruding-mouth retina
Ab34-h3 IHC AB standard conditions Protruding-mouth retinal bipolar neuron nucleus
IHC AB + MO1-znf644a standard conditions Protruding-mouth Not Detected retinal bipolar neuron nucleus
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-ehmt2 standard conditions Long-pec retina ab5-casp3 labeling increased amount, abnormal
Long-pec - Protruding-mouth retina apoptotic process increased occurrence, abnormal
Protruding-mouth retina ab5-casp3 labeling increased amount, abnormal
AB + MO1-ehmt2 + MO4-tp53 standard conditions Long-pec - Protruding-mouth retina apoptotic process process quality, ameliorated
AB + MO1-znf644a standard conditions Long-pec amacrine cell ab1-pax6 labeling absent, abnormal
Long-pec ciliary marginal zone cell vsx2 expression increased distribution, abnormal
Long-pec ciliary marginal zone cell vsx2 expression spatial pattern, abnormal
Long-pec retinal bipolar neuron vsx2 expression increased distribution, abnormal
Long-pec retinal bipolar neuron vsx2 expression spatial pattern, abnormal
Long-pec retinal ganglion cell ab1-pax6 labeling absent, abnormal
Protruding-mouth retinal bipolar neuron ab1-pax6 labeling absent, abnormal
Protruding-mouth retinal bipolar neuron nucleus Ab34-h3 labeling absent, abnormal
AB + MO1-znf644b standard conditions Protruding-mouth retina ab5-casp3 labeling increased amount, abnormal
Protruding-mouth retina apoptotic process increased occurrence, abnormal
AB + MO1-znf644b + MO4-tp53 standard conditions Protruding-mouth retina apoptotic process process quality, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal Stem Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Stem Cell Reports