Fig. 4. SHP-2 deletion promotes myeloid cell differentiation to mature leukocytes with enhanced neutrophil-mediated immunity.

a, ³H-thymidine incorporation (cpm) in OTI splenocytes stimulated with OVA_257–264 cocultured with PMN-MDSC isolated from Shp2^f/f and Shp2^f/fLysM^Cre mice at day 15 post inoculation of MC17-51 tumor cells. Mean ± s.d. of cpm values is shown. ****P < 0.0001 cpm counts obtained by addition of Shp2^f/f versus Shp2^f/fLysM^Cre PMN-MDSC, unpaired t test. Results are representative of three separate experiments using 8–10 mice per group, and three technical replicates per condition. b,c, Quantification (b) and representative histograms (c) of CD38 expression in PMN-MDSC from spleen and BM in mice as in a. Mean percentage ± s.d. d–g, Quantification of MHC II (d) and CD86 (e) expression in tumor-infiltrating CD11b⁺Ly6C^loLy6G⁺ and CD11b⁺Ly6C^hiLy6G⁻ cells and quantification (f) and representative histograms (g) of IFN-γ expression in tumor-infiltrating CD11b⁺Ly6C^loLy6G⁺ and CD11b⁺Ly6C^hiLy6G⁻ cells in mice as in a. Results are from one of three independent experiments with n = 5 mice per group. h,i, Volcano plot of differentially expressed (DE) genes (h) and heat map (i) of top 600 DE genes in PMN-MDSC cells isolated from spleens of Shp2^f/f and Shp2^f/fLysM^Cre tumor-bearing mice and analyzed by bulk RNA-seq (q < 0.05 for all DEGs, log₂(FC) > 0 and log₂(FC) < 0 for upregulated and downregulated genes, respectively). j, Expression of the indicated genes in PMN-MDSC from Shp2^f/f and Shp2^fl/flLysM^Cre tumor-bearing mice (data from RNA-seq dataset). k, Bubble plot of substantially enriched pathways (q < 0.1) in cells from Shp2^fl/flLysM^Cre mice sorted by GeneRatio. l, Heat maps of differentially expressed genes involved in myeloid cell differentiation and neutrophil-mediated immunity (q < 0.05). *P = 0.026, ***P = 0.0034–0.004, ****P < 0.0001, unpaired t test.