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ZFIN ID: ZDB-FIG-161018-25
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: Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Long-pec

Fig. 6

The Retinal Functions of znf644a and znf644b Are Dependent on Functional and Physical Interactions with g9a

(A) Immunostaining of cleaved Caspase3 (red) in retinal cross-sections of embryos injected with individual or combined subphenotypic doses of g9a-MO, znf644a-MO, or znf644b-MO (n = 3 in each group). Yellow arrows highlight apoptotic cells.

(B) (Top) Immunostaining of PCNA (n = 6 in each group) in retinal cross-sections from embryos injected with individual and combined subphenotypic doses of MOs (n = 6 in each group). Yellow arrows denote the position of the PCNA+ population. (Bottom) WISH assays monitoring the expression of vsx2 in retinal cross-sections of embryos injected with individual subphenotypic doses (n = 10 in each group) and combined subphenotypic co-injection of g9a-MO and znf644a-MO (n = 10), g9a-MO and znf644b-MO (n = 13), and znf644a-MO and znf644b-MO (n = 10). Red arrows denote the position of the vsx2 expressing cells.

(C) (Top) WISH assays in retinal cross-sections monitoring vsx2 expression at 48 hpf in znf644a morphant embryos rescued by co-injection of either WT human ZNF644 mRNA (n = 12) or C1263A mutant mRNA (n = 9). (Bottom) Immunostaining of retinal cross-sections monitoring PCNA expression at 48 hpf showing rescue of znf644b morphants by co-injection of WT human ZNF644 mRNA (n = 3) but not C1263A mutants (n = 3).

(D) (Top) WISH assays in retinal cross-sections at 48 hpf monitoring vsx2 expression from znf644b morphant embryos rescued by co-injection of human WT ZNF644 mRNA (n = 14) or C1263A ZNF644 mutant mRNA (n = 9). (Bottom) Immunostaining of retinal cross-sections monitoring cleaved Caspase3 at 72 hpf in znf644b morphant embryos rescued by co-injection of WT human ZNF644 mRNA (n = 3) or a C1263A mutant (n = 3). Red arrows highlight vsx2 expression domains, and yellow arrows highlight PCNA+ cell populations (C) or apoptotic cells (D).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
vsx2 Ab1-vsx2 AB + MO1-ehmt2 + MO1-znf644a standard conditions Long-pec retina central region IHC
Ab1-vsx2 AB + MO1-ehmt2 + MO1-znf644b standard conditions Long-pec retina central region IHC
Ab1-vsx2 AB + MO1-znf644a + MO1-znf644b standard conditions Long-pec retina central region IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab5-casp3 IHC AB + MO1-ehmt2 + MO1-znf644b standard conditions Long-pec retina
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-ehmt2 + MO1-znf644a standard conditions Long-pec retina cell population proliferation increased occurrence, abnormal
Long-pec retina central region vsx2 expression mislocalised, abnormal
AB + MO1-ehmt2 + MO1-znf644b standard conditions Long-pec retina ab5-casp3 labeling increased amount, abnormal
Long-pec retina apoptotic process increased occurrence, abnormal
Long-pec retina cell population proliferation increased occurrence, abnormal
Long-pec retina central region vsx2 expression mislocalised, abnormal
AB + MO1-znf644a + MO1-znf644b standard conditions Long-pec retina cell population proliferation increased occurrence, abnormal
Long-pec retina central region vsx2 expression mislocalised, abnormal
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