Fig. 1

Exposure to E2 Impairs Formation of HSPCs in the AGM

(A) E2 exposure from five somites to 36 hpf decreased runx1/cmyb expression in the AGM (210/234).

(B) E2 diminished runx1:GFP+ cells in the AGM (34/39).

(C) FACS analysis of CD41:GFP embyros (representative samples shown) confirmed that E2 reduced HSCs.

(D) CD41:GFP was significantly decreased by E2 (n = 20, two-tailed t test, p < 0.001; error bars indicate SD).

(E) qPCR quantified reduced runx1 and cmyb expression by E2 (mean of triplicate experiments ± SEM; one-tailed t test runx^∗∗p < 0.01, cmyb^∗p < 0.05).

(F) The pan-Esr antagonist FULV blocked the effect of E2; the isomer 17α-estradiol (10 μM) had no effect (n ≥ 30/treatment).

(G) Estradiol EIA revealed that endogenous estrogens are present during hematopoiesis; E2 was enriched 4.7-fold in the yolk versus body at 18 hpf (mean of triplicate experiments ± SEM; two-tailed t test, ^∗p < 0.05).

(H) ERE:GFP fish show estrogenic activity in the AGM.

(I) GFP expression (as seen in H) was increased 1.57-fold upon E2 exposure (n = 15, two-tailed t test, p < 0.05; error bars indicate SD).

The number of embryos with altered expression over the total number analyzed is shown (bottom right).