PGE₂ and BALF induce LN-like characteristics in BMNs. (A) Bidimensional UMAP analysis of down sampled tissue neutrophils by CD11b, CD62L, CD101, MHC II, Plet1, Sca1 from Zombie NIR fixable viability dye negative adoptive eF670 stained BMNs transferred mice. (B) Heatmap of surface marker expression in transferred eF670⁺ and endogenous eF670⁻ tissue neutrophils. (C) Surface protein expression of CD11b, MHC II, CD14, and CD101 on BMNs following 24 hours of incubation with BALF. (D) Top 20 most highly upregulated genes in LNs compared with BNs by RNA-seq. (E) mRNA expression of Cd14, Tgm2, Nr4a2, and Nr4a3 from vehicle- or BALF-treated BMNs. (F) Surface protein expression of CD14, Plet1, and CD11b following 24 hours of incubation with vehicle, IL-6, or PGE₂. (G) mRNA expression of Tgm2, Cd14, Nr4a2, and Nr4a3 from vehicle-, GM-CSF-, TGF-β-, PGE₂-, or IL-6–treated BMNs (H) GSEA of PGE₂ signaling in LNs compared with BNs. (I) Heatmap of log₂FC value comparing PGE₂- vs vehicle-treated BMNs and LNs vs BNs. (J) Tag density plot of ATAC-seq in PGE₂- (orange) or vehicle- (red) treated BMNs in 107 selected upregulated genes. (K) Representative genome browser snapshot showing signal intensity of ATAC-seq and RNA-seq in annotated sequences in PGE₂- or vehicle-treated BMNs. Arrows indicate the peaks on the promotor regions of each gene. Upper-left number shows the range of read count. (L) Tag counts under ATAC-seq peak spanning the promotor regions of Cd14, Nr4a2, Nr4a3, Tgm2, and G0s2 genes. Data are shown as mean ± standard error of the mean. Significant differences are denoted as *P < .05; **P < .01; ***P < .001; ****P < .0001. % of max scale of each channel is presented as a percentage of maximum count (C,F).