ZFIN Image: Ulloa et al., 2021, Fig. 6

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Figure Caption

Fig. 6

Depletion of HSCs has a negligible impact on T cell seeding of larval thymus

(A) Fluorescent images of drl:CFP-NTR (A′), rag2:mCherry (marking T-lymphocytes) (A′′) and merged (A′′′) at 5 dpf in the thymic lymphoid tissue. Scale bars, 500 μm. Merged fluorescent image (A′′′′) shows higher magnification image of (A′′′).

(B) Experimental schema of the NTR/MTZ system.

(C) Fluorescent images of Tg(drl:CFP-NTR⁺; rag2:mCherry⁺) and Tg(rag2:mCherry⁺) larvae shown at 4 and 6 dpf (2 and 4 dpM, respectively) after depletion of HSCs using the NTR/MTZ system. Yellow arrowheads indicate rag2:mCherry⁺ fluorescent lymphocytes within the thymus. Scale bars, 500 μm.

(D) Quantification of rag2:mCherry fluorescence CTCF levels in Tg(drl:CFP-NTR⁺;rag2:mCherry⁺) and Tg(rag2:mCherry⁺) zebrafish treated with 10 mM MTZ or 1% DMSO (n = 19–72 larvae).

(E) Flow cytometry plots of rag2:mCherry and FITC (autofluorescence control) in untreated negative controls (black); Tg(drl:CFP-NTR;rag2:mCherry) + 1% DMSO (purple); Tg(rag2:mCherry) + 10 mM MTZ (light blue); and Tg(drl:CFP-NTR;rag2:mCherry) + 10 mM MTZ (red).

(F) Quantification of rag2:mCherry% from (E) flow cytometry experiments in treated and control groups; n = 4–11, 7–10 pooled larvae per sample.

Two-way ANOVA with Tukey’s multiple comparisons test was used for all statistical analyses. Plots are individual data points for each biological replicate with mean ± SEM. ns, not significant; *p < 0.05; **p ≤ 0.01; ***p ≤ 0.001.

Acknowledgments

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