Figure 3.

Global GPCR Analysis in the AGM

(A) Global GPCR analysis.

(B) Non-olfactory GPCRs (N = 314) were categorized into Secretin, Adhesion, Glutamate, Frizzled/Taste2, and Rhodopsin families based on sequence homology.

(C) Classification of 85 GPCRs expressed in the AGM (>5 transcripts per million) into five families.

(D) Bar graph depicting GATA-2-regulated genes from RNA-seq analysis of +9.5^+/+ and +9.5^−/− AGMs (Gao et al., 2013). Black bars, genes co-regulated by GATA-1 according to our prior microarray analysis of G1E-ER-GATA with or without β-estradiol treatment (DeVilbiss et al., 2013).

(E) Gata2, Adora3, Gpr65, Ltb4r1, and P2ry1 expression during erythropoiesis. B, basophilic erythroblast; O, polyorthochromatic erythroblast; P, proerythroblast; R, reticulocyte (http://www.cbil.upenn.edu/ErythronDB/).

(F) Time course of Gata2 and Gpr65 expression following β-estradiol treatment in G1E-ER-GATA cells (n = 3 independent experiments).

(G) RT-PCR analysis of Gata2 and Gpr65 in FACS-sorted R1, R2, R3, and R4/5 populations from fetal liver (n = 3 independent experiments).

(H and I) ChIP signal map for Gpr65 in human CD34 cells (H) (Beck et al., 2013), mouse HPC7 cells (Wilson et al., 2010), Lin⁻ bone marrow cells (Li et al., 2011), and G1E cells (Trompouki et al., 2011) (I).

(J and K) RT-PCR analysis of Gata2 and Gpr65 mRNA in +9.5^+/+ and +9.5^−/− AGM (5 litters: +9.5^+/+ [n = 8 embryos]; +9.5^−/− [n = 6 embryos]) and yolk sac (three litters: +9.5^+/+ [n = 7 embryos]; +9.5^−/− [n = 5 embryos]) (J), and MAE cells expressing GATA-2 (K) (n = 3 independent experiments).

(L) RNA-seq of Gata2 and Gpr65 mRNA in FACS-sorted endothelial cells (EC), hemogenic endothelial cells (HEC), hematopoietic cells (HC), and hematopoietic stem cells (HSC) from the AGM (Solaimani Kartalaei et al., 2015).

Error bars represent SEM. ^∗p < 0.05; ^∗∗∗p < 0.001 (two-tailed unpaired Student's t test).