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. 2020 Mar 24;4(6):1145–1158. doi: 10.1182/bloodadvances.2019000785

Figure 6.

Figure 6.

Dysregulation of TLR4 signaling pathway in Runx1 KO neutrophils occurs prior to the committed neutrophil stage. (A) Representative FACS plots of intracellular TNF-α production by neutrophils (CD11b+Ly6G+) after stimulation of whole BM with vehicle or 100 ng/mL LPS for 4 hours. Runx1 PMN KO = Runx1f/f; MRP8-Cre. (B) Quantification of the frequency of TNF-α+ neutrophils and relative MFI of the TNF-α+ neutrophils normalized to control neutrophils run in the same experiment (n = 4 from 4 experiments, mean ± SD, 1-way analysis of variance and Tukey’s multiple comparisons test). (C) Presence of the expected wild-type, floxed (f), or deleted Runx1 polymerase chain reaction products from tail clips (with a small amount of contaminating blood) and FACS-purified neutrophils (CD11b+SiglecF−F4/80−Ly6G+). (D) Western blot showing total RUNX1 levels in FACS-purified neutrophils and splenocytes from the same mice. ***P ≤ .001; ****P ≤ .0001. ns, not significant.