Journal: PLOS One

Article Title: The combination of LILRB4-targeting NK cell engagers and cGAS–STING agonists enhances the anti–multiple myeloma immune activity of NK cells

doi: 10.1371/journal.pone.0339375

Figure Lengend Snippet: (A) Uniform manifold approximation and projection (UMAP) dimensionality reduction clustering identifies multiple cell subpopulations of multiple myeloma cells. (B) The Volcano plots display the gene expression profiles of each tumor cell subpopulation with significant differences compared to other subpopulations. Log2 (fold change) > 1 and adjusted P < 0.05 were considered as significantly differentially expressed genes. (C) Heatmap showing precise differences in LILRB4 expression across tumor cell subsets. (D) Density plot of LILRB4 expression in tumor cells from UMAP. Color intensity indicates the expression level. (E) Copy Number Variation Score for Tumor Subpopulation Cells. (F) Metabolic pathway enrichment analysis of differentially expressed genes in tumor cell subsets, where color intensity reflects the degree of correlation. (G) Dotplot diagrams display the expression of ligand–receptor pairs between LILRB4 + MM cells (the initiating point of communication) and other cells. (H) LILRB4-related survival analysis in MM was performed using the TCGA database, with P values calculated by the log-rank test. (I) Western blot analysis was used to detect LILRB4 protein expression levels in MM1.S and RPMI-8226 cells. (J) Flow cytometry was employed to assess the expression of LILRB4 protein in MM1.S and RPMI-8226 cells.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 2 hours, incubated overnight at 4°C with primary antibodies (1:1000 dilution, rabbit anti-human, Proteintech), Rabbit anti-human LILRB4 antibody was used to detect the expression of LILRB4 in MM cell lines, which used GAPDH as an internal reference.

Techniques: Gene Expression, Expressing, Western Blot, Flow Cytometry

Journal: PLOS One

Article Title: The combination of LILRB4-targeting NK cell engagers and cGAS–STING agonists enhances the anti–multiple myeloma immune activity of NK cells

doi: 10.1371/journal.pone.0339375

Figure Lengend Snippet: (A) Schematic representation of the BiKE structure. (B) Schematic representation of BiKE-mediated NK cells killing of tumors. (C) BiKE SDS-PAGE of non-reduced and reduced proteins, stained with Coomassie blue. (D) ELISA test measuring the affinity of different concentrations of BiKE to the LILRB4 antigen. (E) Elisa tests the affinity of different concentrations of BiKE to the CD16A antigen. (F) Indirect labeling Flow cytometry was used to assess the binding ability of BiKE to MM1.S, NK, and K562 cells. Goat anti human IgG (H + L) antibody labeled with FITC as the secondary antibody. (G) PBMCs were co-cultured with LILRB4 + MM1.S cells for 4 hours. The regulation of CD69 expression in NK cells by monoclonal antibodies and BiKE in the co-culture system was analyzed by flow cytometry. Data are presented as mean ± SD, *P < 0.05, ***P < 0.001, ****P < 0.0001, n = 4 compared to each other group by one-way ANOVA.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 2 hours, incubated overnight at 4°C with primary antibodies (1:1000 dilution, rabbit anti-human, Proteintech), Rabbit anti-human LILRB4 antibody was used to detect the expression of LILRB4 in MM cell lines, which used GAPDH as an internal reference.

Techniques: SDS Page, Staining, Enzyme-linked Immunosorbent Assay, Labeling, Flow Cytometry, Binding Assay, Cell Culture, Expressing, Bioprocessing, Co-Culture Assay

Journal: Clinical and Experimental Immunology

Article Title: Early exposure of interferon-? inhibits signal transducer and activator of transcription-6 signalling and nuclear factor ?B activation in a short-term monocyte-derived dendritic cell culture promoting 'FAST' regulatory dendritic cells

doi: 10.1111/j.1365-2249.2011.04537.x

Figure Lengend Snippet: Interferon (IFN)-γ in combination with the FAST-dendritic cell (DC) protocol generates phenotypically tolerogenic DC. Monocytes were isolated from peripheral blood and cultured without (UT-DC) or with IFN-γ (IFN-γ-DC) in the presence of interleukin (IL)-4 and granulocyte–macrophage colony-stimulating factor (GM-CSF) for 24 h. DC were then either matured with tumour necrosis factor (TNF)-α (10 ng/ml) and prostaglandin E2 (PGE2) (1 µM) for another 24 h (a: plus TNF-α and PGE2, representative of six independent experiments) or were cultured for another 24 h in the absence (b: minus TNF-α and PGE2, representative of three independent experiments. Fluorescence activated cell sorter (FACS) analysis was used to determine phenotypic markers. The following co-stimulatory and linage markers were examined: CD83, CD80, CD86, immunoglobulin (Ig)-like transcript 4 (ILT4), CD11c, DC-SIGN, CD14, human leucocyte antigen D-related (HLA-DR). Line indicates the isotype control and solid histogram shows the tested monoclonal antibody. Negative gate was set to <0·5%.

Article Snippet: Antibodies The phenotypic profile of DC was defined using the following directly conjugated monoclonal antibodies (mAb): anti-CD83-fluorescein isothiocyanate (FITC) (HB15e), anti-CD86-FITC (FUN1), anti-CD80-FITC (L307·4), anti-DC-SIGN-FITC (DCN46), anti-human leucocyte antigen D-related (HLA-DR)-phycoerythrin-cyanin-5 (PE-Cy5) (G46-6) (BD Bioscience, San Jose, CA, USA) and rat anti-human immunoglobulin (Ig)-like transcript 4 (ILT4) (42D1) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used as a primary mAb and FITC-conjugated anti-rat IgG was used for detection.

Techniques: Isolation, Cell Culture, Fluorescence, Control

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: Elevated LILRB4 expression predicts poor prognosis in OC. (A) Expression profiles of LILRB family genes in OC cohort from TCGA database. (B) Comparison of LILRB4 mRNA expression levels between OC and normal tissues in the TCGA dataset combined with GTEx database. (C–H) Validation of LILRB4 mRNA expression differences between OC and normal tissues across 6 independent GEO datasets ( GSE12470 , GSE18520 , GSE40595 , GSE46169 , GSE51927 , and GSE66957 ). (I) Further validation of LILRB4 expression in human OC and normal tissues collected from Shanghai First Maternity and Infant Hospital. (J) Protein expression levels of LILRB4 in OC tissues (T) and adjacent non-tumor tissues (P) were assessed by WB (left). Densitometric quantification was performed using ImageJ software and normalized to GAPDH expression (n = 5) (right). (K) IHC staining showing LILRB4 expression in tumor and adjacent non-tumor tissues. Scale bar = 200 μm. (L) Kaplan–Meier survival analysis based on TCGA and GEO datasets evaluating the association between LILRB4 expression and OS and PFS. Statistical significance was determined using the log-rank test. (M) ROC curve analysis showing the diagnostic performance of LILRB4, with the AUC indicating predictive accuracy. Data are presented as mean ± standard deviation (SD). Statistical significance was determined using an unpaired two-tailed Student’s t-test. *p < 0.05, **p < 0.01, ****p < 0.0001.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Expressing, Comparison, Biomarker Discovery, Software, Immunohistochemistry, Diagnostic Assay, Standard Deviation, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: Downregulation of LILRB4 suppresses the malignant phenotypes of OC cells. (A) mRNA expression levels of LILRB4 in the normal ovarian epithelial cell line IOSE80 and multiple OC cell lines (HEY, A2780, CAOV3, OVCAR8, and ES2). (B) WB analysis confirming the protein knockdown efficiency of LILRB4 in OVCAR8 (left) and ES2 (right) cells; representative images are shown. (C) WB analysis confirming the protein overexpression efficiency of LILRB4 in HEY; representative images are shown. (D, E) Cell proliferation of OVCAR8 (D) and ES2 (E) cells in the NC and LILRB4 knockdown groups, as assessed by CCK-8 assay. (F) Proliferation of HEY cells with NC or LILRB4 overexpression, evaluated by CCK-8 assay. (G–I) Transwell assays demonstrating the migration (up) and invasion (down) abilities of OVCAR8 (G) , ES2 (H) and HEY (I) cells under the indicated treatments. All images are shown at the same magnification with a scale bar of 100 μm. (J) WB analysis confirming the protein knockdown efficiency of Lilrb4 in the ID8 cell line; representative images are shown. (K) Representative images of orthotopic tumors in C57BL/6 mice (n = 4 per group) implanted with ID8 cells (NC vs. sh-Lilrb4), with a scale bar of 10 mm. (L) Comparison of tumor weight (up) and tumor volume (down) between the two groups shown in (K) . (M) IHC analysis of Ki67 expression in orthotopic tumor tissues, including representative images (left) and quantification of Ki67-positive areas (right) (n = 3). All panels are shown at the same magnification with a scale bar of 100 μm. Data are presented as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test (D, E, G, H), or unpaired two-tailed Student’s t-test (F, I, L, M). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Expressing, Knockdown, Over Expression, CCK-8 Assay, Migration, Comparison, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: LILRB4 is associated with macrophages in the OC immune microenvironment. (A) Correlation of StromalScore, ImmuneScore, and ESTIMATEScore with LILRB4 expression in TCGA-OV patients. (B) UMAP visualization of single-cell RNA-seq data from the OV_EMTAB8107 tumor dataset (left) and corresponding LILRB4 expression across different cell clusters (right). (C) Distribution of LILRB4 expression across various cell types in GEO-derived single-cell datasets, analyzed using the TISCH2 database ( http://tisch.comp-genomics.org/ ), with darker colors indicating higher expression levels. (D) Correlation between macrophage enrichment scores and LILRB4 expression in TCGA-OV samples, as determined by ssGSEA. (E) Relative mRNA expression of F4/80 in orthotopic ovarian tumor models (NC and sh-LILRB4 groups). (F) Representative IHC images showing F4/80 expression in orthotopic ovarian tumors (n = 3). Scale bar, 100 μm. (G) Representative IHC staining of CD68 and LILRB4 in corresponding regions of human OC tissues (n = 3). All panels are shown at the same magnification. Scale bar, 100 μm. Data are presented as mean ± SD. Statistical analysis was performed using unpaired two-tailed Student’s t-test. *p < 0.05, ****p < 0.0001.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Expressing, Single Cell, RNA Sequencing, Derivative Assay, Immunohistochemistry, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: LILRB4 regulates M2c macrophage polarization. (A) Patients in the TCGA-OV cohort were stratified into high- and low-LILRB4 expression groups to assess differences in M2 macrophage infiltration. (B) LILRB4 expression levels in M0, M1, and M2 macrophages based on the GEPIA database. (C) TCGA-OV samples were categorized into high and low M2 macrophage infiltration groups. OS was analyzed based on LILRB4 expression using the TIMER database; statistical significance was determined by the log-rank test. (D) Correlation analysis LILRB4 and CD206 (M2a), HLA-DR (M2b), CD163 (M2c), CD14 (M2d) by GSE12470 cohort. (E) Correlation analysis between LILRB4 expression and the M2c markers CD163 in the TCGA-OV cohort. (F, H) Flow cytometry analysis of CD163 expression in macrophages following co-culture with OVCAR8 (F) or ES2 (H) cells with NC or LILRB4 knockdown. (G, I) qRT-PCR analysis of IL-10 and TGFβ1 mRNA expression in macrophages co-cultured with OVCAR8 (G) or ES2 (I) cells under control (NC) or LILRB4 knockdown conditions. (J) Flow cytometry analysis of CD163 expression in macrophages following co-culture with HEY cells with NC or OE-LILRB4. (K) A schematic diagram showing the process of collecting tumor cell CM and co-culturing it with macrophages for 48 hours before conducting flow cytometry detection. (L) Macrophages were treated with CM from LILRB4-overexpressing tumor cells or control tumor cells, and the proportion of CD163-positive macrophages was assessed (left) and quantitatively compared between the 2 groups (right). (M) Macrophages were treated with CM from LILRB4-overexpressing tumor cells or control tumor cells, and the expression levels of IL-10 and TGFβ1 in macrophages were measured by qRT-PCR. Data are presented as mean ± SD. Statistical analysis was performed using unpaired two-tailed Student’s t-test (A, F-J, L, M). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Expressing, Flow Cytometry, Co-Culture Assay, Knockdown, Quantitative RT-PCR, Cell Culture, Control, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: LILRB4 promotes M2c macrophage differentiation via CCL5/NF-κB signaling. (A) In the OVCAR8 cells, qRT-PCR was used to analyze the mRNA expression levels of CCL2, CCL5, CCL18, CCL22, CXCL12 and CXCL8. (B) Correlation analysis between CCL5 (left) and CCL22 (right) expression and M2 macrophage infiltration based on the TIMER database. (C) Expression levels of the M2c macrophage marker CD163 in macrophages following treatment with CCL5 or CCL22. (D) The expression of CCL5 mRNA in ES2 cells either under normal control conditions or under sh-LILRB4 treatment was analyzed by qRT-PCR. (E) The expression of CCL5 mRNA in HEY cells under different treatment was analyzed by qRT-PCR. (F) qRT-PCR analysis of CCL5 mRNA expression in ovarian tumor tissues from the indicated groups. (G) Correlation analysis between LILRB4 expression and CCL5 in the TCGA-OV cohort. (H) Flow cytometry analysis of CD11b + CD163 + macrophages in the control and CCL5-treated groups (left), with corresponding quantification shown on the right. (I) Correlation analysis between CCL5 expression and key signaling molecules (NFKB1, AKT1, mTOR, STAT6, MAPK1, and NOTCH1) in the TCGA-OV cohort. (J) WB analysis showing p-p65 levels in macrophages treated with CM from control or LILRB4-overexpressing tumor cells or maraviroc, indicating activation of the NF-κB signaling pathway. (K) Expression levels of CD163 in macrophages under the above treatment conditions (up), with quantitative analysis of the different groups (down). (L) Assessment of p-p65 activation in macrophages following CCL5 treatment. (M) qRT-PCR analysis of CD163, IL-10, and TGFβ1 mRNA expression in macrophages treated with control (NC) or CCL5 or CCL5 with PDTC. Data are presented as mean ± SD. Statistical analysis was performed with unpaired two-tailed Student’s t-test (A, C-F, H) or unpaired two-tailed Student’s t-test (K, M). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Quantitative RT-PCR, Expressing, Marker, Control, Flow Cytometry, Activation Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: Clinical significance of LILRB4 expression in OC. (A–B) Differences in LILRB4 expression levels were compared among subgroups with venous invasion (A) and lymphatic invasion (B) in the TCGA-OV cohort. (C–D) Correlation analysis between LILRB4 and immune checkpoint molecules in the TCGA-OV cohort, presented as a chord diagram (C) and a lollipop plot (D) . (E) Kaplan–Meier survival analysis of patients from the immunotherapy cohort (esophageal adenocarcinoma treated with immune checkpoint inhibitor-based therapy). OS (left) and PFS (right) were compared between tumors with high and low LILRB4 expression. Patients were stratified according to the optimal cutoff determined by the Kaplan–Meier Plotter platform. Statistical significance was assessed using the log-rank test. (F) Representative images of orthotopic ID8 tumors in C57BL/6 mice (n = 4 per group). (G) Comparison of tumor weight (left) and volume (right) among different groups of C57BL/6 mice (n = 4 per group). (H) IHC analysis of Ki-67 and CD163 expression in tumor tissues (n = 3). All images were captured at the same magnification; scale bar = 100 μm. (I–J) WB analysis of p-p65 expression in tumor tissues (I, n = 3) and corresponding quantitative analysis (J) . (K) IHC analysis of CD8 and GZMB expression in tumor tissues (n = 3). All images were captured at the same magnification; scale bar = 100 μm. (L) A nomogram was constructed based on independent prognostic factors for overall survival. Data are presented as mean ± SD. Statistical analysis was performed using an unpaired two-tailed Student’s t-test. *p < 0.05, **p < 0.01, ****p < 0.0001.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Expressing, Comparison, Construct, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: Graphic abstract of LILRB4 in modulating the TIME in OC. An increase in LILRB4 expression promotes tumor development and acts as a key regulator of immune remodeling. It enhances CCL5 secretion, thereby activating NF-κB signaling in macrophages and promoting M2c macrophage polarization. This immunosuppressive reprogramming facilitates tumor growth and progression within the TME. Therefore, targeting LILRB4 may represent a promising therapeutic strategy to improve immunotherapy efficacy, enhance antitumor immunity, and achieve better clinical outcomes in OC.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Expressing