Decreased production of HPCs in the absence of Gpr56 and Gpr97. (A) Schematic of Gpr56 and Gpr97 deletion in mouse Gpr56 locus. (B) RT-PCR analysis of time course hematopoietic differentiation cultures of G2V WT and G2V.56^KO97^KO ESCs. Relative Gpr97 expression (normalized to β-actin). (C) Representative flow cytometry plots for CD45⁺ hematopoietic cells and EMPs (CD41⁺cKit⁺CD16/32⁺) from day 10 differentiated G2V WT, G2V.56^KO, and G2V.56^KO97^KO ESC cultures. Percentages shown in each quadrant. (D) Percentage of viable cells and fold change in percentages of V⁺, CD45⁺, and EMPs from day 10 differentiated G2V WT, G2V.56^KO, and G2V.56^KO97^KO ESC cultures. n = 3. Mean ± SEM. (E) Frequency of CFU-C in day 10 differentiated G2V WT, G2V.56^KO, and G2V.56^KO97^KO ESC cultures. CFU-C per 10⁴ V⁺ cells is shown with colony types indicated by color. Mean ± SEM. (F) Output of CFU-C per 10⁴ V⁺ in the different colony types: BFU-erythroid, CFU- E, -G, -M, -GM, -GEMM. *P ≤ .05; **P ≤ .01; ***P ≤ .001.