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. 2022 Sep 29;185(20):3720–3738.e13. doi: 10.1016/j.cell.2022.08.018

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© 2022 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Figure S1 — mTOR deficiency impairs hematopoiesis in zebrafish, related to Figure 2

(A) Overlaid widefield fluorescence and bright-field micrographs of an mtor^fh178/fh178 animal and wild-type sibling expressing the neutrophil-specific fluorescent reporter Tg(lysC:EGFP) 6 dpf (Hall et al., 2007).

(B) Numbers of neutrophils in the caudal hematopoietic tissue (CHT) of animals from mtor^fh178/+; Tg(lysC:EGFP) incross 2 and 6 dpf.

(C–F) Zebrafish embryos were manually dechorionated and treated with 400 nM rapamycin or 0.5% DMSO on 1 dpf to block primitive and intermediate waves of hematopoiesis (Clements and Traver, 2013).

(C) Confocal micrographs of the CHT of Tg(cd41:GFP) zebrafish at 2 dpf. Hematopoietic stem cells (HSCs, open arrowheads) and thrombocytes, nucleated counterparts of platelets in non-mammalian vertebrates, (filled arrowheads) were identified by low versus high level expression of cd41:GFP, respectively (Lin et al., 2005; Ma et al., 2011).

(D) Numbers of HSCs in the CHT of 2 dpf animals.

(E) Numbers of thrombocytes in the CHT of 2 dpf animals.

(F) Numbers of macrophages in the midbrain and CHT of Tg(mpeg1:YFP) zebrafish 2 dpf.

Scale bars: 300 μm in (A) and 100 μm in (C). (B and D–F) Symbols represent individual animals. Horizontal lines indicate means. Statistical analyses, (B) two-way ANOVA with Tukey’s post-test and (D–F) unpaired Student’s t test.