Fig 2: Overall survival (OS) according to PD-L1 tumor expression (50% cutoff); n: 3,289.

Fig 3: Knockdown of NPM1 suppresses PD-L1 expression and promotes T cell activity in vivo.a 0.5 × 106 4T1 cells were injected into the mammary fat pads of BALB/c mice. The average tumor volume of each group was depicted over time. b The number of lung metastasis nodes of every mouse in each group (n = 8) was graphed, and the metastasis rate in teach group was indicated above the bar. c Tumor infiltrating lymphocytes (TILs) in the tumors of each group (n = 5) were analyzed by flow cytometry. d Granzyme B and CD8 in mouse tumor tissues of each group were detected by immunofluorescence (IF). Experiment was repeated in three animal samples with similar results. e NPM1 and PD-L1 expression was detected by IHC in mouse tumor tissues of each group. Experiment was repeated in three animal samples with similar results. Data were presented as mean ± s.d. of three independent experiments. a–c were analyzed by one-way ANOVA + two-side Tukey test. *P < 0.05, **P < 0.01, ****P < 0.0001; ns, not significantly different. Source data are provided as a Source Data file.

Fig 4: NPM1 binds to PD-L1 promoter and upregulates PD-L1 expression in TNBC cells.a Fragments of PD-L1 promoter in the luciferase reporter plasmids. b Activity of the different fragments of PD-L1 promoter was determined by dual-luciferase assay. n = 3 independent experiments. c Schematic diagram of the streptavidin-agarose pull-down assay. d Proteins from the pull-down assay were separated by polyacrylamide gel eletrophoresis and revealed by sliver staining. The protein band marked in red rectangle was cut out to be analyzed by mass spectrometry. Three biological replicate samples were used. e Binding of NPM1 to PD-L1 promoter was examined by ChIP-qPCR. n = 3 independent experiments. f PD-L1 promoter activity in MDA-MB-231 cells transiently transfected with control siRNA (nc), NPM1 siRNAs (si-NPM1-1, si-NPM1-2), control plasmid for NPM1(vector), or NPM1 overexpression plasmid (NPM1 oe) was measured by dual-luciferase assay. g, h PD-L1 mRNA levels (g), NPM1 and PD-L1 protein expression (h) in MDA-MB-231 and HCC1806 cells with transient NPM1 knockdown or overexpression was detected. i Jurkat cells were co-cultured with MDA-MB-231 or HCC1806 cells in a 1:4 ratio for 48 h, and IL-2 secreted was detected by ELISA. j Primary T cells were co-cultured with MDA-MB-231 and HCC-1806 cells in a 1:30 ratio for 24 h. IFN-? producing T cells were detected by flow cytometry. k Western blot analysis of NPM1 and PD-L1 expression in MDA-MB-231 cells with NPM1 knockdown and/or PD-L1 overexpression. l, m MDA-MB-231 cells with NPM1 knockdown and/or PD-L1 overexpression were co-cultured with Jurkat cells (l) or primary T cells (m). Data were presented as mean ± s.d. of three independent experiments. f Right panel, g 2th and 4th panel were analyzed by two-side Student’s t-test, **P < 0.01, ***P < 0.001, ****P < 0.0001. f Left panel, g 1th and 3th panel, i, j were analyzed by one-way ANOVA + two-side Dunnett test, **P < 0.01, ***P < 0.001, ****P < 0.0001. l, m were analyzed by one-way ANOVA + two-side Tukey test, **P < 0.01, ****P < 0.0001; ns not significantly different. Source data are provided as a Source Data file.

Fig 5: Overall survival (OS) according to PD-L1 tumor expression (1% cutoff); n: 4,262.