Fig 2: Kaplan-Meier overall survival curves of PD1/PDL1 expression or CD8+T cells/CD68+M density in GC. GC patients with MSI, PD1 positive, and PDL1[T]/PDL1 negative had better prognosis (P=0.006, 0.012, 0.005 and 0.022, respectively). There are no correlations between CD8+T cells/CD68+M density and prognosis in GC (P=0.870 and 0.985, respectively).

Fig 3: MTHFD2 is induced by IFN-γ and involved in IFN-γ-mediated PD-L1 regulation.a, b Indicated cancer cells transfected with control or MTHFD2 siRNAs for 24 h and then treated with 0 or 20 ng/ml IFN-γ for another 24 h, PD-L1 mRNA levels were analyzed by real-time PCR (a) and immunoblotting analyses were performed using the indicated antibodies (b). c, d Indicated cancer cells pretreated with LY294002 (c) and Rapamycin (d) with indicated concentrations and then cultured with 20 ng/ml IFN-γ, immunoblotting analyses were performed using the indicated antibodies. e Indicated cancer cells treated with 20 ng/ml IFN-γ for indicated times, immunoblotting analyses were performed using the indicated antibodies. In (a), the values are presented as mean ± s.e.m (n = 3 independent experiments); p values (Student’s t test, two-sided) with control or the indicated groups are presented (also see Supplementary Fig. 3).

Fig 4: Expression of immune‐checkpoint molecule (ICM) ligands in head and neck squamous cell carcinoma (HNSCC) tissues. ICM ligands expression in the nest of case 11 is shown (A). Representative double immunostaining images with programmed cell death‐1 (PD‐1) in orange and cytokeratin in blue (upper right), galectin 9 in red and cytokeratin in blue (lower left) and CECAM‐1 in green and cytokeratin in blue (lower right) are shown in the same area with HE images (upper left). ICM ligand expression in the stroma is shown (B). Representative double immunostaining images with PD‐L1 in orange and CD204 in blue (upper right), CD68 in green and CD204 in blue (lower left) and galectin‐9 in red and CD204 in blue (lower right) are shown in the same area with the HE images (upper left). Distribution of programmed cell death receptor ligand 1 (PD‐L1) (orange) expressing macrophages along the periphery of the nest (cytokeratin in blue) was observed (C, upper panel) in an “immune‐excluded” case (case 14). In an “immune‐inflamed” case (case 24), high infiltration of PD‐L1 (orange)‐expressing macrophages into the nest (cytokeratin in blue) were observed (D, upper panel). These macrophages were positive for CD204 (C and D, lower panel). Upper right and lower right images of (C) and (D) indicate the expanded images of the white square left of the images. Magnifications are indicated in each image

Fig 5: RT-PCR analysis of effect of miR-191-5p on PD-L1 in colon cancer lines. (A) RT-PCR analysis of the expression level of miR-191-5p in colon cell lines and NCM460 cells. (B and C). RT- PCR analysis of the expression levels of miR-191-5p and PD-L1 after the transfection with miR-191-5p inhibitor in LoVo cells or miR-191-5p mimic in RKO cells. ** P < 0.001, * P < 0.05.