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. Author manuscript; available in PMC: 2024 Oct 3.
Published in final edited form as: Cell Metab. 2023 Oct 3;35(10):1688–1703.e10. doi: 10.1016/j.cmet.2023.09.004

Figure 4. Tumor-secreted GM-CSF induces Acod1 in neutrophils through the STAT5-C/EBPβ axis.

Figure 4.

(A) ER-Hoxb8-immortalized mouse myeloid progenitors proliferate in the presence of estrogen (β-estradiol) and stem cell factor (SCF). When estrogen is withdrawn, progenitors differentiate to mature neutrophils (ER-Hoxb8-DNs) and can be induced to a TIN-like status with tumor CM or specific cytokines. (B) Western blot to examine Acod1 expression by ER-Hoxb8-DNs induced with CM from murine mammary cancer cell lines. (C) Western blot of Acod1 and C/EBPβ (LAP) in ER-Hoxb8 progenitor cells (first lane) and ER-Hoxb8-DNs (other lanes) treated with 4T1-expressed cytokines individually (unit ng/ml) or 4T1 CM. (D) t-SNE plot of Acod1 and Cebpb from the Drop-seq data. (E) Western blot of Acod1 and C/EBPβ (LAP) for ER-Hoxb8-DNs with Cebpb knockdown by shRNA (three designs) and induced with GM-CSF. (F) Western blot of Acod1 and C/EBPβ (LAP) for E0771-CM-treated BMDNs with or without αGM-CSF. (G) Western blot of Acod1 and C/EBPβ (LAP) for ER-Hoxb8-DNs induced with GM-CSF in the presence of BP-1–102 (STAT3 inhibitor) or STAT5-IN-1 (STAT5 inhibitor). (H) Schematic of Acod1 upregulation in TINs by tumor-secreted GM-CSF. Western blot results were representative of at least three independent experiments showing consistent patterns. See also Figure S4.