|
Fig. 1 The commensal microbiota is required for HSPC development (A) CONR and GF zebrafish were examined by WISH for cmyb expression in the caudal hematopoietic tissue (CHT) region at 5 and 6 dpf. Arrowheads denote cmyb+ HSPCs. (B) Quantification of WISH results in (A). (C) qPCR analysis of runx1 and cmyb expression in the kidneys of CONR and GF zebrafish at 7 dpf. (D) WISH for neutrophil marker (mpx), leukocyte marker (L-plastin), T lymphocyte marker (rag1), and erythrocyte marker (hbbe1) at 5 dpf in CONR and GF zebrafish. (E) Quantification of WISH results in (D). (F) The experimental timeline for the conventionalization (CONV) experiments in (G–J). (G) WISH for cmyb and mpx in CONR, GF, and CONV fish in the CHT region at 5 dpf. (H) Quantification of WISH results in (G). (I) WISH for cmyb in the kidney in CONR, GF, and CONV fish at 7 dpf. (J) qPCR analysis of runx1 and cmyb expression in the kidneys of CONR, GF, and CONV zebrafish at 7 dpf. Student’s t tests in (B, C, and E), ANOVA in (H, I, and J); mean ± SEM from more than three independent experiments, n = 45–60 larvae per group for WISH, 30–40 larvae per group for qPCR, ∗p < 0.05, ∗∗p < 0.01; ns, not significant. CONR, conventionally raised; GF, germ free; CONV, conventionalized. Scale bars, 200 μm. See also Figures S1 and S2.