Fig. 2: Identification of IL36G-producing CiMs.

a Schematic of sample collection for bulk RNA-Seq (n = 2 independent samples per each). b Principal component analysis for bulk RNA-seq data of BM CD115⁺ Ly6C^high inflammatory monocytes obtained from the indicated mice. c HOPACH-generated clusters of differentially expressed genes in CD115⁺ Ly6C^high inflammatory monocytes from the indicated mice. d Heat map of re-clustered genes upregulated in Mo-Leukemia mice (HOPACH algorithm) with statistically enriched Gene Ontology (GO) terms (left) and enriched GO term-associated genes (right). The upregulated genes in Mo-Leukemia mice (c) were re-clustered. e, f Receptor-ligand-related genes in DEGs upregulated in CD115⁺ Ly6C^high inflammatory monocytes from cachectic Mo-Leukemia mice (e). For the six selected genes, volcano plots in the indicated comparisons are shown in (f). g, h Plasma membrane-related genes among DEGs upregulated in CD115⁺ Ly6C^high inflammatory monocytes from cachectic Mo-Leukemia mice (g). For the selected 95 genes, a volcano plot in the indicated comparison is shown in (h). i, j Flow cytometric analysis of total PB from the control, Mo-Leukemia mice with cachexia, and Mo-Leukemia mice without weight loss (n = 5 mice per group). Representative flow cytometry plots (i) and the frequencies of CD38⁺ cells in PB Ly6C⁺ inflammatory monocytes (j) are shown. k Diff-Quik staining of CD38⁺ Ly6C⁺ inflammatory monocytes from Mo-Leukemia mice with cachexia. Scale bar, 10 μm. l Flow cytometric analysis of the total BM from control, Mo-Leukemia mice with cachexia, and Mo-Leukemia mice without weight loss (n = 4 mice per each). The frequencies of CD38⁺ cells in BM Ly6C⁺ inflammatory monocytes are shown. m Relative diameters of C2C12 myotubes after the co-culture assay with the indicated cells. Representative results from more than three independent experiments are shown. The number of muscle fibers analyzed in each sample is shown. n Schematic of sample collection. o mRNA expression levels of Il36g in control Ly6C⁺ monocytes (n = 4 mice), CD38⁺ Ly6C⁺ inflammatory monocytes from Mo-Leukemia mice (n = 5 mice), and CD38^‒ Ly6C⁺ inflammatory monocytes from Mo-Leukemia mice (n = 5 mice). p Expression level of Il36g protein in the culture supernatant of control Ly6C⁺ monocytes (n = 4 mice), CD38⁺ Ly6C⁺ inflammatory monocytes from Mo-Leukemia mice (n = 5 mice), and CD38^‒ Ly6C⁺ inflammatory monocytes from Mo-Leukemia mice (n = 5 mice). Statistical analyses were performed using Fisher’s exact test (d), empirical Bayes moderated t test (f, h), and one-way ANOVA followed by Tukey multiple comparison test (j, l, m, o, p). Data are presented as the mean ± SD (j, l, m, o, p). Source data are provided as a Source Data file.