FIGURE

Fig. 3

ID

ZDB-FIG-220701-51

Publication

Basheer et al., 2022 - In vivo impact of JAK3 A573V mutation revealed using zebrafish

Other Figures

All Figure Page

Back to All Figure Page

Fig. 3

Effect of Jak3 A573V mutation on later hematopoiesis. A, B Gene expression analysis. Total RNA extracted from 28 dpf homozygous wild-type (jak3^+/+), heterozygote (jak3^+/ca) and homozygote (jak3^ca/ca) mutant zebrafish was subjected to qRT²PCR analysis (A) with gene markers of hematopoietic stem cells (cmyb), T cells (rag1, cd4, cd8), B cells (cd79a, igm), and NK cells (nccrp1, nkld). Data are represented as relative fold change compared to homozygous wild-type (+/+) fish, with mean and SEM shown in red and statistical significance compared for Cq values normalized to control actb indicated (*p < 0.05; n = 3), or to RT-PCR (B) with primers specific for T cell receptor (TCR) β-chain (v(d)jcβ vb1.5, vb12, vb14.5) and B cell Ig heavy chain (igVH1, igVH4) rearrangements and actb as the control and analyzed by agarose gel electrophoresis. RT-negative controls yielded no products (data not shown) (n = 2). C–J Histological analysis. Giemsa stained blood cells (C–E) and kidney cells (G–I) of adult wild-type (jak3^+/+) (C, G), heterozygote (jak3^+/ca) (D, H) and homozygote (jak3^ca/ca) (E, I) fish, with differential counts for blood (F) and kidney (J) performed (e: erythrocyte, eo: eosinophil, l: lymphocyte, n: neutrophil, p: precursor, m: macrophage) (n = 4–8; 100 × magnification; scale bar = 10 μm). K–P FACS analysis. Kidney cells from adult wild-type (jak3^+/+) (K), heterozygote (jak3^+/ca) (L) and homozygote (jak3^ca/ca) (M) were subjected to FACS analysis, with lymphoid (N) myeloid (O) and precursor (P) populations quantified, with mean and SEM shown in red and level of statistical significance indicated (*p < 0.05, **p < 0.01; n = 5). Q Survival analysis. Wild-type (jak3^+/+), heterozygote (jak3^+/ca) and homozygote (jak3^ca/ca) zebrafish were assessed weekly for survival, which is displayed as a Kaplan–Meier plot with level of statistical significance indicated (***p < 0.001, n = 35). R Gene expression analysis. Total RNA extracted from adult homozygous wild-type (jak3^+/+), heterozygote (jak3^+/ca) and homozygote (jak3^ca/ca) zebrafish was subjected to qRT²PCR analysis with gene markers of hematopoietic stem cells (cmyb), T cells (tcra), B cells (cd79a), and NK cells (nccrp1), as described for panel A

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Cell. Mol. Life Sci.