Machine learning-based prediction reveals kinase MAP4K4 regulates neutrophil differentiation through phosphorylating apoptosis-related proteins
Fig 4
Loss of MAP4K4 expression impairs the differentiation process of bone marrow neutrophils.
(A) Overview of study design, created using Biorender. (B) UMAP Plots for single-cell gene expression pooled across bone marrow samples, clusters visualized and labeled by cell type. The plot is split by control (Ctrl) vs Map4k4-cKO (cKO). (C) Proportions of the six neutrophil clusters in control (Ctrl) and Map4k4-cKO (cKO) samples. The p value was calculated by the adjusted proportion test. (D) Numbers of neutrophil progenitor cells in the bone marrow of control (Ctrl) or Map4k4-cKO (cKO) mice; c-KithiLy6Gneg (myeloblasts, MB); c-KitintLy6Gneg (promyelocytes, PM); c-KitnegLy6Glow (myelocytes, MC); c-KitnegLy6Gint (metamyelocytes, MM); and c-KitnegLy6Ghi (band cells and segmented neutrophils, BC/SC); BM, bone marrow (Ctrl n=7, cKO n=6; mean ± SD). Mann-Whitney U test. (E) ROS generation of neutrophil progenitor cells in BM of control (Ctrl) or Map4k4-cKO (cKO) mice; MFI, mean fluorescent intensity; ROS, reactive oxygen species; MB, myeloblasts; PM, promyelocytes; MC, myelocytes; MM, metamyelocytes; BC/SC, band cells and segmented neutrophils; (n=6; mean ± SD). Mann-Whitney U test. (F) The number of DEGs in control (Ctrl) vs Map4k4-cKO (cKO) of six neutrophil subpopulations. (G) GO-BP analysis of cluster-based DEGs between control (Ctrl) and Map4k4-cKO (cKO) HSC to neutrophils. Selected GO terms with Benjamini-Hochberg-corrected p values < 0.05 (one-sided Fisher’s exact test) are shown. The dot size represented the number of genes. The color scale represented the adjusted p value. Fig 4A was created using Biorender.