Additional file 2 of Tumor cell-imposed iron restriction drives immunosuppressive polarization of tumor-associated macrophages

Additional file 2: Figure S2. Tumor cells fail to educate macrophages toward an iron-releasing phenotype but instead an iron starvation response. (A) The mRNA expression of Tfrc, Fth1, and SLC40A1 in PMs co-cultured with indicated cells or DMEM alone using qPCR. The fold changes in expression level relative to control were expressed as 2−ΔΔCt. (B) Heat map of iron metabolism-related genes in THP-1 monocyte-derived macrophages based on RNA-seq data from GSE159254. (C) BMDMs and Hepa1-6 cells in the co-culture system were allowed to grow in DMEM media containing 50 ng/mL FITC-labeled holo-transferrin, and then in vitro holo-transferrin uptake assay was performed. BMDMs were collected 2 h later, and the MFI of FITC-transferrin was measured by FACS. All data are representative of three independent experiments and presented as mean ± SD. *p < 0.05, ***p < 0.001