ZFIN ID: ZDB-FIG-161220-4
Iwanami et al., 2016 - Forward Genetic Screens in Zebrafish Identify Pre-mRNA-Processing Pathways Regulating Early T Cell Development. Cell Reports   17:2259-2270 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Day 5
PHENOTYPE:
Fish:
Observed In:
Stage Range: Protruding-mouth to Day 5

Fig. 5

Characterization of tnpo3 Mutants, Related to Figure S5

(A) Reduction of ikaros-expressing cells in the thymus (encircled with dotted lines in middle and right panels) of homozygous tnpo3 mutants (additionally transgenic for an ikaros:eGFP reporter). Note the normal numbers of ikaros-positive cells in general hematopoietic tissues at 1 dpf. Scale bars, 100 μm.

(B) Maturation block of thymocytes. Few ccr9b-positive thymocytes (insets in 4 dpf panels) in tnpo3 mutants express rag1. Scale bars, 100 μm.

(C) Thymopoietic indices at 5 dpf. Each symbol represents one animal. The mean ± SEM is indicated. Scale bar, 100 μm.

(D) Analysis of ptprc isoforms by RT-PCR; size markers are indicated.

(E) Masking genetic interaction between gemin5 and tnpo3 mutations in zebrafish. The thymopoietic indices (rag1/gh ratio; mean values ± SD) at 5 dpf are given for fish (genotypes denoted in bars) resulting from crosses of parents heterozygous for both gemin5 and tnpo3; the observed thymopoietic index for gemin5−/−;tnpo3−/− double mutants is significantly higher than that expected from the indices of single mutants under the multiplicative model of genetic interaction. The numbers of fish in each group is indicated.

(F) Alternative splicing events in tnpo3 mutants at 4 dpf. Bar chart shows the number of significant AS events per condition at a false discovery rate (FDR) of 1% for five different AS event types. The total numbers of tested AS events are indicated on the right.

(G) Genes affected by exon-skipping events.

(H) Genes exhibiting transcriptional upregulation.

(I) Genes exhibiting transcriptional downregulation.

(J) Summary of the network structure involving snapc3, lsm8, gemin5, and tnpo3 genes. +, alleviating genetic interaction; −, synthetic interaction; ?, unclear.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ccr9b WT standard conditions Day 4 thymus ISH
tnpo3t22074/t22074 standard conditions Day 4 thymus ISH
EGFP fr101Tg standard conditions Prim-5 hematopoietic system IFL
Protruding-mouth thymus IFL
Day 5 thymus IFL
tnpo3t22074/t22074 ; fr101Tg standard conditions Prim-5 hematopoietic system IFL
Protruding-mouth thymus IFL
Day 5 thymus IFL
gh1 WT standard conditions Day 5 hypophysis ISH
tnpo3t22074/t22074 standard conditions Day 5 hypophysis ISH
rag1 WT standard conditions Protruding-mouth thymus ISH
Protruding-mouth thymus T cell ISH
Day 4 thymus ISH
Day 4 thymus T cell ISH
Day 5 thymus ISH
tnpo3t22074/t22074 standard conditions Protruding-mouth thymus ISH
Protruding-mouth thymus T cell ISH
Day 4 thymus ISH
Day 4 thymus T cell ISH
Day 5 thymus ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
gemin5t26393/t26393 standard conditions Day 5 T cell differentiation decreased occurrence, abnormal
gemin5t26393/t26393 ; tnpo3t22074/t22074 standard conditions Day 5 T cell differentiation decreased occurrence, abnormal
tnpo3t22074/t22074 standard conditions Protruding-mouth T cell differentiation decreased occurrence, abnormal
Protruding-mouth thymus rag1 expression decreased amount, abnormal
Protruding-mouth thymus T cell decreased amount, abnormal
Day 4 mRNA processing decreased occurrence, abnormal
Day 4 mRNA processing process quality, abnormal
Day 4 T cell differentiation decreased occurrence, abnormal
Day 4 thymus rag1 expression decreased amount, abnormal
Day 4 thymus T cell decreased amount, abnormal
Day 5 T cell differentiation decreased occurrence, abnormal
Day 5 thymus rag1 expression decreased amount, abnormal
tnpo3t22074/t22074 ; fr101Tg standard conditions Protruding-mouth T cell differentiation decreased occurrence, abnormal
Protruding-mouth thymus EGFP expression decreased amount, abnormal
Protruding-mouth thymus T cell decreased amount, abnormal
Day 5 T cell differentiation decreased occurrence, abnormal
Day 5 thymus EGFP expression decreased amount, abnormal
Day 5 thymus T cell decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.