Reduced RhoA-GTPase activity in cKO P2 neutrophils leads to enhanced speed in highly confined environments. (A) The protein-protein interaction (PPI) network map was built from the 49 genes from GSEA signatures: “cytoskeleton remodeling” and the small GTPases. Bold lines are potential PPIs between RhoA or Cdc42 and GSEA signature gene products, and thin lines associates both signature genes. Genes in yellow are defined as master regulators, and genes in brown contain a putative HIF2α binding site (supplemental Figure 4A,C). (B-D) Rho GTPase activity measured in steady-state BMDNs from all different genotypes and their WT littermates. Data points represent individual mice from ≥2 experiments - normalized values against WT control. Data are mean ± standard error of the mean. Quantification of average speed of WT neutrophils treated with different inhibitors against RhoA (E) or Cdc42 inhibitor (F) in 3-µm-wide microchannels. Data are represented as box plots (+median), and whiskers range from the 10th percentile to the 90th percentile. Data points are representative of 2 independent experiments. *P < .05, Mann-Whitney U test.