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Additional file 1: Figure S1. Characterization of circRanGAP1 in HCC. Figure S2. circRanGAP1 expression in HCC cells. Figure S3. CircRanGAP1 acted as a sponge of miR-27b-3p in HCC cells. Figure S4. miR-27b-3p plays tumor-suppressive roles in HCC. Figure S5. Figure S6. The effect of NRAS in cirRanGAP1 knockdown cells. Figure S7. Immune infiltration analysis of miR-27b-3p and NRAS. Figure S8. The effects of circRanGAP1 on immunocytes infiltration. Table S1. Sequences of Primers used for qRT-PCR. Table S2. List of Primary Antibodies Used in the Study. Table S3. Target sequences of hsa_circ_0063513 shRNAs. Table S4. circ_0063513 and miR-27b-3p FISH probe sequences. Table S5. The potential targets of RanGAP1.

Additional file 1: Table S1. The functional enrichment analysis of 100 VIPR1-related genes. Figure S1. Immune infiltration analysis of VIP and ADCYAP1 A. C. Explored the correlation between immune infiltration and VIP and ADCYAP1 through the ssGSEA algorithm in the “GSVA” R package. B. D. Evaluated the correlation between Treg/Tem/NK/iDC cells and VIP, Tem/NK/iDC/Mast cells and ADCYAP by the Pearson correlation analysis. *P

Additional file 4: Table S1. List of primers used for qPCR analysis in this study.

Additional file 1: Figure S1. Iron deprivation drives the immunosuppressive polarization of macrophages. (A) Cell viability was measured in BMDMs and PMs with indicated treatment by CCK-8 assay. (B) The mRNA expression of M1 signature genes in PMs with indicated treatment using qPCR. The fold changes in expression level relative to control were expressed as 2−ΔΔCt. (C) The mRNA expression of M2 signature genes in RAW264.7 cells with indicated treatment using qPCR. The fold changes in expression level relative to control were expressed as 2−ΔΔCt. (D–G) The surface expression of CD206 and PD-L1 with indicated treatment in RAW264.7 cells using FACS. The results were shown as relative fold changes in MFI of CD206 and PD-L1 normalized to their corresponding controls. All data are representative of three independent experiments and presented as mean ± SD. *p

Additional file 3: Figure S3. Iron deprivation drives macrophage M2 polarization through induction of Hif-1α. (A, B) The mRNA expression of Hif-1α and Hif-2α in PMs with indicated treatment using qPCR. The fold changes in expression level relative to control were expressed as 2−ΔΔCt. (C) The protein expression of HIF-2α in PMs with indicated treatment was detected by Western blot. (D) The mRNA expression of Arg1, Vegfa, and Pd-l1 by qPCR in BMDMs with either Hif-2α knockdown or its control counterpart upon indicated treatment. The fold changes in expression level relative to control were expressed as 2−ΔΔCt. (E) The concentration of lactate in the supernatants of Hepa1-6/shNT and Hepa1-6/shTfrc cells was measured by lactate assay kit.

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