Aberrant CD14 overexpression on granulocytes was functionally significant in the pathogenesis of del(5q) MDS. (A) The levels of serum TNFα after intraperitoneal injection of LPS with different doses into indicated mice. (B) Real-time polymerase chain reaction (PCR) analysis of TNFα and IL-6 mRNA levels in Gr1/Mac1 double-positive granulocytes treated with LPS for different times. (C) Western blot analysis of phospho–inhibitory nuclear factor κB (IκB) α and total IκBα levels in cells from panel B at indicated time points. Heat shock cognate protein 70 (Hsc70) was used as a loading control. (D) Real-time PCR analysis of IκBα mRNA level in cells from panel B at indicated time points. (E) Injection of high-dose LPS (30 µg/g) into indicated mice (6-8 weeks old, whole body knockout). The fraction of survival over time was plotted using Kaplan-Meier survival analysis. N = 11 in each group. Data were from 2 independent experiments. (F) Same as in panel E except that WT mice 1 month after transplantation of total bone marrow cells from the indicated mDia1 mice were used. N = 8 in each group. (G) CBC for neutrophils, red blood cells, and platelets 6 months after weekly intraperitoneal injection of LPS (2 µg/g). N = 6 in each genotype group. Mice were 6-8 weeks old at the time of first injection. (H) Wright-Giemsa stains of bone marrow smears (top panels) and H&E stains of bone marrow sections (lower panels) of indicated mice from panel F. Arrows indicate dysplastic granulocytes. Scale bars represent 10 μm (upper) and 50 μm (lower).