Dysregulated expression of inflammatory pathways in Runx1 KO neutrophils. (A-D) RNA-seq data collected from FACS-purified neutrophils (CD11b⁺SiglecF⁻F4/80⁻Ly6G⁺) stimulated with vehicle or 100 ng/mL LPS for 2 hours (n = 3 from 3 experiments). Expression values are normalized to total cell number with a spike in control. (A) GO analysis of differentially expressed genes upregulated in Runx1 KO neutrophils as compared with control neutrophils with vehicle treatment. GO analysis of differentially expressed genes downregulated in vehicle-treated Runx1 KO neutrophils as compared with controls and up- or downregulated in LPS-treated Runx1 KO neutrophils as compared with controls is shown in supplemental Figure 6A. (B) Heat maps of TLR4 pathway genes in vehicle-treated Runx1 KO (RV1, RV2, RV3) compared with control neutrophils (CV1, CV2, CV3) ordered by fold-change in expression (high to low). Statistically significantly upregulated genes are marked with an asterisk and green text (false discovery rate <0.05). (C) Normalized expression of TLR4 pathway genes in vehicle-treated neutrophils (P = 1.45e-5, Student t test) (D) Schematic of TLR4 pathway genes with all statistically significantly upregulated genes in vehicle-treated Runx1 KO neutrophils denoted in green. Schematic adapted from O’Neill et al.⁴⁵  (E) Cell surface TLR4 on control and Runx1 KO neutrophils (CD11b⁺Ly6G⁺) as compared with a representative isotype control (normalized to mode). Bar graphs depict absolute TLR4 MFI of individual samples (n = 3, mean ± SD, 2-tailed unpaired Student t test). Data are representative of 3 experiments. **P ≤ .001.