Analysis of PU.1 expression in the BM and spleen of WT and Hbb^th3/+ mice. Representative histograms of fluorescence intensity of PU.1 expression in neutrophils (Ly6G^highLy6C^low), immature myeloid cells (Ly6G^lowLy6C^low), and monocytes (Ly6G^negLy6C^high) in (A-C) BM and (D-F) spleen from WT and Hbb^th3/+ mice. Yellow shading represents isotype control; blue shading represents WT mice, and red shading represents Hbb^th3/+ mice. PU.1 protein expression levels determined using flow cytometric analysis in (G) neutrophils (Ly6G^highLy6C^low), (H) immature myeloid cells (Ly6G^lowLy6C^low), and (I) monocytes (Ly6G^negLy6C^high) from the BM and spleen of WT and Hbb^th3/+ mice. ΔMFIs were calculated by subtracting isotype MFIs from PU.1 MFIs. Symbols represent results from individual animals. Statistical significance between groups was determined using Student t test. RT-qPCR analysis of PU.1 mRNA expression levels relative to housekeeping gene GAPDH in sorted (J) neutrophils (Ly6G^highLy6C^low), (K) immature myeloid cells (Ly6G^lowLy6C^low), and (L) monocytes (Ly6G^negLy6C^high) from the BM and spleen of WT and Hbb^th3/+ mice. Data are presented as mean ± SEM, n ≥ 5 per mice group. The statistical significance between the groups was evaluated with Student t test.