recombinant mouse interferon γ ifn γ  (R&D Systems)

Journal: EMBO Reports

Article Title: Human ZBP1 is a potent inducer of cell death through mechanisms divergent from mouse ZBP1

doi: 10.1038/s44319-026-00866-6

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Recombinant Mouse IFN-γ , ABclonal , Cat#RP01070.

Techniques: Recombinant, Sequencing, Magnetic Beads, Protease Inhibitor, Software, Microscopy, Bicinchoninic Acid Protein Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: IFN-γ suppresses tumor growth and invasion. (A) Cytokine profiling of co-culture supernatants via ELISAs: IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α. (B-D) Spatial expression patterns of IFN-γ. (B) Immunofluorescence imaging of the invasive front in SSIT, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green), and grayscale intensity distribution. (C) Immunofluorescence imaging of TIM and NIM, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green). (D) Quantification of relative IFN-γ expression in TIM and NIM. (E) Representative Ki-67 immunohistochemistry images of SSIT cases stratified into IFN-γ-high and IFN-γ-low groups (n=5 each; median split). (F) Quantification of Ki-67 index comparing the two groups. (G) EdU staining demonstrating dose-dependent suppression of TtT/GF pituitary adenoma cell proliferation by IFN-γ (0–100 ng/ml; 48 h). (H) Representative flow cytometry histograms for cell cycle analysis of cells treated with IFN-γ (0–100 ng/ml) in the absence (0 µM) or presence (5 µM) of ruxolitinib. (I) Stacked bar plot showing the percentages of cells in the G 1 , S and G 2 /M phases under the same treatment conditions. (A) One-way ANOVA with Tukey's post hoc multiple comparisons test. (D and F) Unpaired two-tailed Student's t-test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. CTRL, control; DMC, digested mucosal culture; EdU, 5-ethynyl-2′-deoxyuridine; IBA-1, ionised calcium binding adaptor molecule 1; MTC, mucosal tissue culture; NIM, non-invaded mucosa; ns, not significant; PE-A, phycoerythrin-area; SSIT, sphenoid sinus-invasive tumor; TIM, tumor-invaded mucosa.

Article Snippet: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

Techniques: Co-Culture Assay, Expressing, Immunofluorescence, Imaging, Immunohistochemistry, Staining, Flow Cytometry, Cell Cycle Assay, Two Tailed Test, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Elevated IgG levels drive macrophage M2-to-M1 reprogramming. (A) Sphenoid sinus-invasive tumor cases stratified into CD19-high (n=5) and CD19-low (n=5) groups based on the cohort median of CD19 + B cell density, with (B) quantitative analyses of macrophage polarization (M1-like versus M2-like). (C) Dural-invasive tumor and non-invasive tumor cases stratified into IgG-high (n=27) and IgG-low (n=26) groups based on the cohort median of relative IgG immunohistochemistry staining intensity, with (D) quantitative analyses of M1-like/M2-like macrophage proportions. (E and F) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by IgG (10 µg/ml) exposure. Relative (E) IL-6 and (F) TNF-α mRNA expression in RAW264.7 macrophages pre-polarized to M0, M1 or M2 states. (G) Representative flow cytometric cell-cycle profiles of TtT/GF cells following the indicated treatments. (H) Stacked bar plot summarizing the percentages of cells from (G) in G 1 , S and G 2 /M phases. (I) Representative images from the scratch wound assay at 0, 24, 48 and 72 h under the indicated treatments. (J) Quantification of scratch wound closure. (K) Representative western blot images showing total STAT1, p-STAT1, total STAT3, p-STAT3 and β-actin levels in cells treated with IFN-γ (100 ng/ml), IL-6 (100 ng/ml), IFN-γ + IL-6 (50 ng/ml each), ruxolitinib (5 µM) or IFN-γ + IL-6 (50 ng/ml each) plus ruxolitinib (5 µM), as indicated. (L) Densitometric semi-quantification of p-STAT1/STAT1 (ratio). (B and D) Unpaired two-tailed Student's t-test. (E, F, J and L) One-way ANOVA with Tukey's post hoc multiple comparisons test. *P<0.05, ***P<0.001, ****P<0.0001. CTRL, control; IBA-1, ionised calcium binding adaptor molecule 1; ns, not significant; p-, phosphorylated; PE-A, phycoerythrin-area.

Article Snippet: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

Techniques: Immunohistochemistry, Staining, Expressing, Scratch Wound Assay Assay, Western Blot, Two Tailed Test, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Anti-CD47 mAb enhances ADCP to suppress tumor cell proliferation. (A) Immunofluorescence staining of CD47 (red) and DAPI (blue) in a representative subset of non-invasive tumor, dural-invasive tumor and sphenoid sinus-invasive tumor cases (n=10 per group). (B) Paired comparison of CD47 fluorescence intensity at the IF versus the TC. (C) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by anti-CD47 mAb (10 µg/ml) treatment for 12 h. Quantitative PCR was used to analyze polarization/activation markers. (D) Schematic illustrating anti-CD47 mAb-mediated blockade of the CD47-SIRPα axis and enhancement of ADCP. (E) EdU assay of TtT/GF cell proliferation in a Transwell co-culture with anti-CD47 mAb-treated polarized macrophages. (F) Quantification of EdU-positive cells. (G) Representative microscopy images and flow cytometry plots showing macrophage phagocytosis of pHrodo™ Red-labeled GFP-TtT/GF cells. (H) Quantification of phagocytosis (%). (B) Paired two-tailed Student's t-test. (C, F and H) One-way ANOVA with Tukey's post hoc multiple comparisons test. **P<0.01, ***P<0.001, ****P<0.0001. ADCP, antibody-dependent cellular phagocytosis; Arg-1, arginase 1; EdU, 5-ethynyl-2′-deoxyuridine; FcγR, Fcγ receptor; GFP, green fluorescent protein; IF, invasive front; mAb, monoclonal antibody; NOS2, nitric oxide synthase 2; ns, not significant; PE, phycoerythrin; SIRPα, signal regulatory protein-α; SSCA, side scatter area; TC, tumor core.

Article Snippet: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

Techniques: Immunofluorescence, Staining, Comparison, Fluorescence, Real-time Polymerase Chain Reaction, Activation Assay, EdU Assay, Co-Culture Assay, Microscopy, Flow Cytometry, Labeling, Two Tailed Test

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Summary graphic illustration. This illustration summarizes the proposed model during pituitary adenoma invasion. The tumor invasive front abuts an intact sphenoid sinus mucosa, forming a distinct boundary. The mucosal compartment is enriched for ionised calcium binding adaptor molecule 1-positive macrophages with an M1-like predominance and IgG-high B cells. B cell-derived IgG promotes M2-to-M1 macrophage reprogramming, while coordinated IFN-γ and IL-6 production establishes a tumor-suppressive cytokine gradient that decreases from mucosa toward the tumor core, constraining proliferation and migration via JAK-STAT1 activation. Therapeutically, anti-CD47 monoclonal antibody blocks the CD47-SIRPα ‘don't-eat-me’ axis and augments antibody-dependent cellular phagocytosis, highlighting a strategy for immune checkpoint-targeted therapy that may complement surgical management. FcR, Fc receptor; JAK, Janus kinase; mAb, monoclonal antibody; p-, phosphorylated; SIRPα, signal regulatory protein-α.

Article Snippet: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

Techniques: Binding Assay, Derivative Assay, Migration, Activation Assay

Journal: Redox Biology

Article Title: A novel photosensitizer-based photodynamic therapy reprograms the Kynurenine–AhR axis to boost antitumor immunity in breast cancer

doi: 10.1016/j.redox.2026.104171

Figure Lengend Snippet: DTP-PDT attenuates the IDO–Kyn–AhR axis and relieves immune-suppression in the TME. (A-D) Targeted metabolomics analysis of Trp, Kyn, QA, and 5-HT in cell samples. (E) Targeted metabolomics of Kyn in tumor tissue samples. (F) Levels of Kyn in culture supernatants after the indicated various treatments under IFN-γ priming (n = 3). (G) Representative immunofluorescence images of tumor sections stained for CD3 (green), AhR (red), and DAPI (blue). Scale bar = 20 μm. (H) RT-qPCR analysis of Cyp1a1, Cyp1b1, and Ahrr mRNA expression in tumor-infiltrating CD3 + T cells (n = 4). (I-J) Proportion of intratumoral CD8 + T cells (gated on CD3 + T cells, n = 5). (K-L) Proportion of intratumoral Treg cells (gated on CD3 + CD4 + Foxp3 + T cells, n = 5). (M) Representative immunofluorescence images of tumor sections stained for CD3 (green), AhR (red), and DAPI (blue) in the Kyn rescue experiment. Scale bar = 20 μm. (N) RT-qPCR analysis of Cyp1a1, Cyp1b1, and Ahrr mRNA expression in tumor-infiltrating CD3 + T cells from the Kyn rescue experiment (n = 4). (O–P) Proportion of intratumoral CD8 + T cells in the Kyn rescue experiment (gated on CD3 + T cells, n = 5). (Q-R) Proportion of intratumoral Treg cells in the Kyn rescue experiment (gated on CD3 + CD4 + Foxp3 + T cells, n = 5). Data are shown as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: 4T1 cells (5 × 10 5 cells/well) were seeded in 6-well plates and allowed to adhere for 24 h. To activate IDO1 activity, cells were pretreated with recombinant mouse interferon-γ (IFN-γ) (50 ng/mL, 485-MI, R&D System) for 24 h. After IFN-γ priming, cells were treated with DTP-PDT and/or the IDO1 inhibitor (NLG919, Aladdin).

Techniques: Immunofluorescence, Staining, Quantitative RT-PCR, Expressing

Journal: Bioactive Materials

Article Title: Carrier free oral Co-delivery of atorvastatin via baicalein-copper-network for atherosclerosis therapy through senescence reversal and multi-mechanistic synergy

doi: 10.1016/j.bioactmat.2025.12.036

Figure Lengend Snippet: Reactive oxygen species scavenging capacity of CMA. (A) H 2 O 2 (100 μM) scavenging capacity CMA at 2 h. (B) O 2 − scavenging capacity of CMA at 40 min. (C) •OH scavenging capacity of CMA at 1 min. (D) DPPH (100 μg mL -1 ) scavenging capacity of CMA at 1 h. (E) Fluorescence images of RAW264.7 cells treated with CMA and LPS + IFN-γ for 24 h. Intracellular ROS were stained with DCFH-DA (Green), and nuclei were stained with DAPI (blue). Scale bar = 50 μm. (F) Quantitative analysis of mean DCFH-DA fluorescence intensity. Flow cytometric analysis of ROS expression in: (G) RAW264.7: CMA + LPS; (H) RAW264.7: CMA; (I) SMC: CMA; (J) HUVEC: CMA; (K) RAW264.7: CMA + BAPTA; (L) SMC: CMA + BAPTA; (M) HUVEC: CMA + BAPTA. All treatments were conducted for 24 h. Data are presented as mean ± SD (n = 3–6; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001).

Article Snippet: Baicalein (BAI), copper chloride (CuCl 2 ·2H 2 O), and Atorvastatin (ATV) were purchased from Macklin Inc. Lipopolysaccharides (LPS), recombinant mouse interferon γ (IFN-γ), oxidized low-density lipoprotein (oxLDL), dihydroethidium (DHE), DiI-oxidized low-density lipoprotein (DiI-oxLDL), hematoxylin-eosin (H & E) stain kit, modified Masson's trichrome stain kit, and modified Oil Red O stain kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. Cy5-baicalein was purchased from Xi'an Qiyue Biology.

Techniques: Fluorescence, Staining, Expressing

Journal: Bioactive Materials

Article Title: Carrier free oral Co-delivery of atorvastatin via baicalein-copper-network for atherosclerosis therapy through senescence reversal and multi-mechanistic synergy

doi: 10.1016/j.bioactmat.2025.12.036

Figure Lengend Snippet: Macrophage reprogramming ability of CMA. (A) Representative optical images of RAW264.7. Scale bar = 50 μm. (B) Confocal laser scanning microscopy images of RAW264.7 cells stained with CD206 antibody, with nuclei counterstained with DAPI. Scale bar = 50 μm. (C) Quantitative analysis of mean CD206 fluorescence intensity. (D) Flow cytometric analysis of CD206 expression in RAW264.7 treated with BAI, Cu-PBS, Cu-MON, ATV, and CMA for 24 h. (E) Relative expression levels of Arg-1, VEGF, TNF-α, and IL-1β in cell supernatants. (F) Flow cytometry scatter plots of iNOS and CD206 expression in RAW264.7 pretreated with LPS + IFN-γ for 24 h followed by treatment with BAI, Cu-PBS, Cu-MON, ATV, and CMA for 24 h. (G–I) Flow cytometric analysis of M1/M2 expression (MFI ratio), iNOs expression, CD206 expression in RAW264.7. (J) Relative expression levels of TGF-β, Arg-1, VEGF, TNF-α, and IL-1β in supernatants from cells treated as in F. Data represent mean ± SD (n = 3–6 independent experiments). Statistical significance: ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001 versus Control; ns = not significant.

Article Snippet: Baicalein (BAI), copper chloride (CuCl 2 ·2H 2 O), and Atorvastatin (ATV) were purchased from Macklin Inc. Lipopolysaccharides (LPS), recombinant mouse interferon γ (IFN-γ), oxidized low-density lipoprotein (oxLDL), dihydroethidium (DHE), DiI-oxidized low-density lipoprotein (DiI-oxLDL), hematoxylin-eosin (H & E) stain kit, modified Masson's trichrome stain kit, and modified Oil Red O stain kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. Cy5-baicalein was purchased from Xi'an Qiyue Biology.

Techniques: Confocal Laser Scanning Microscopy, Staining, Fluorescence, Expressing, Flow Cytometry, Control

Journal: Cell Reports Medicine

Article Title: HE4 drives PD-L1 expression in myeloid cells via IFN-γR-JAK-STAT3 signaling to promote tumor immune evasion

doi: 10.1016/j.xcrm.2026.102691

Figure Lengend Snippet: HE4 promotes tumor immune evasion by upregulating PD-L1 expression within TME (A–C) scRNA-seq analysis of two paired tumor and paratumor samples from LUAD patients showing cell clustering (A), WFDC2 /HE4-expressing cell populations (B), and quantification of WFDC2 /HE4 expression in epithelial/malignant clusters (C). (D) Online dataset analysis of WFDC2 mRNA expression in tumor versus normal LUAD tissues. (E and F) HE4 overexpression (E) or knockout (F) respectively promoted or suppressed the growth of subcutaneous LLC tumors in mice. (G) Survival of mice intraperitoneally inoculated with HE4-overexpressing or control ID8 cells. (H) HE4 knockout attenuated ID8 peritoneal tumor progression, shown by representative abdominal images and ascites volume. (I) SDS-PAGE with Coomassie blue staining showing the purity of Fc and mouse HE4-Fc (mHE4-Fc) recombinant proteins. (J) Administration of mHE4-Fc promoted MC38 subcutaneous tumor growth. (K) mHE4-Fc administration reversed the growth suppression of HE4-KO LLC subcutaneous tumors. (L) mHE4-Fc failed to reverse tumor growth suppression in HE4-KO LLC tumors implanted in Rag1 -deficient mice. (M) Extracellular HE4 did not promote LLC cell proliferation in vitro , as assessed by CCK8 assay. (N) mHE4-Fc administration suppressed CD8 + T cell activation in the TME of HE4-KO LLC tumors, assessed by IFN-γ + and granzyme B + CD8 + T cells. (O) HE4 upregulated PD-L1 expression on macrophages in the microenvironment of LLC, MC38, and HE4-KO LLC tumors. (P–R) mHE4-Fc administration failed to reverse tumor suppression of HE4-KO LLC subcutaneous tumors in Cd274 -deficient mice, with treatment scheme (P), tumor growth (Q), and tumor weight (R). Statistical analyses were performed using Wilcoxon rank-sum test (D), two-way ANOVA (E, F, J–L, and Q), log rank (Mantel-Cox) test (G), and two-tailed paired (C) or unpaired t tests (H, M, N, O, and R). Data represent two independent experiments (E and M).

Article Snippet: Recombinant Fc-tagged mouse IFN-γ protein , MedChemExpress , Cat# HY- P73252.

Techniques: Expressing, Over Expression, Knock-Out, Control, SDS Page, Staining, Recombinant, In Vitro, CCK-8 Assay, Activation Assay, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: HE4 drives PD-L1 expression in myeloid cells via IFN-γR-JAK-STAT3 signaling to promote tumor immune evasion

doi: 10.1016/j.xcrm.2026.102691

Figure Lengend Snippet: HE4 competes with IFN-γ for IFN-γR binding and modulates downstream gene expression (A and B) Raw264.7 cells were stimulated with HE4-Fc (20 μg/mL) or IFN-γ (100 ng/mL) for 3 h, followed by RNA-seq; volcano plots of HE4- (A) or IFN-γ-regulated genes (B) are shown. (C) Genes commonly upregulated by HE4 and IFN-γ. (D) AlphaFold-3-predicted interfaces of HE4-IFNGR1/2 and IFN-γ-IFNGR1/2 complexes, with shared receptor-contact residues highlighted. (E) Competitive binding assay: His-tagged IFNGR1/2 was incubated with Flag-HE4 in the presence or absence of IFN-γ, followed by Ni-TED pull-down and immunoblotting. (F and G) SPR sensorgrams showing binding of mHE4-Fc (F) or mIFN-γ-Fc (G) to mIFNGR1-His. (H and I) ELISA quantification of HE4 and/or IFN-γ levels in ascites from ID8-tumor-bearing mice (H) and LLC-tumor-conditioned media (I). (J) High concentrations of HE4 reduced IFN-γ binding to IFNGR1/2 in competitive pull-down assays. (K) PCA of RNA-seq profiles from Raw264.7 cells treated with HE4-Fc, HE4-Fc plus IFN-γ, or IFN-γ for 12 h. (L) Expression (TPM) of representative STAT1- or STAT3-associated genes following the indicated treatments. Statistical analyses were performed using two-tailed paired Student’s t tests (H and I). Data in (E) and SPR sensorgrams (F and G) are representative of three independent experiments.

Article Snippet: Recombinant Fc-tagged mouse IFN-γ protein , MedChemExpress , Cat# HY- P73252.

Techniques: Binding Assay, Gene Expression, RNA Sequencing, Competitive Binding Assay, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: HE4 drives PD-L1 expression in myeloid cells via IFN-γR-JAK-STAT3 signaling to promote tumor immune evasion

doi: 10.1016/j.xcrm.2026.102691

Figure Lengend Snippet: HE4 blockade promotes antitumor immunity in the tumor microenvironment (A–C) scRNA-seq analysis of LLC tumors from mice treated with control IgG or anti-HE4 antibody ( n = 3 per group), showing UMAP clustering of 26 cell populations (A), relative abundance of each cluster (B), and aggregated cell types (C). ∗ p < 0.05 (D and E) HE4 neutralization reduced Cd274 (PD-L1) expression in myeloid compartments. UMAP feature plots show Cd274 expression in macrophage/monocyte and epithelial/malignant populations (D), with paired comparison across macrophage clusters (E). (F) UMAP visualization of nine intratumoral T cell subclusters in control IgG– and anti-HE4–treated tumors. (G) In the LLC subcutaneous model, intratumoral IFN-γ + and CD69 + CD8 + T cells were quantified by flow cytometry. (H) In the ID8 intraperitoneal model, IFN-γ + CD8 + T cells were quantified by flow cytometry. (I–K) HE4 neutralization failed to suppress LLC tumor growth in Rag1 −/− mice, shown by treatment scheme and tumor growth/endpoint analyses. (L–O) CD8 + T cell depletion abrogated the antitumor efficacy of HE4 blockade, with treatment scheme, tumor growth/endpoint measurements, and confirmation of depletion efficiency by flow cytometry. (P–S) Macrophage depletion using anti-CSF1R diminished the antitumor efficacy of HE4 neutralization, with tumor growth/endpoint measurements and confirmation of depletion efficiency by flow cytometry. Statistical analyses were performed using unpaired t tests (B, C, H, and K), paired t test (E), one-way ANOVA (G, N, O, R, and S), and two-way ANOVA (J, M, and Q). Data in (G, H, and L–S) are pooled from two independent experiments.

Article Snippet: Recombinant Fc-tagged mouse IFN-γ protein , MedChemExpress , Cat# HY- P73252.

Techniques: Control, Neutralization, Expressing, Comparison, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: HE4 drives PD-L1 expression in myeloid cells via IFN-γR-JAK-STAT3 signaling to promote tumor immune evasion

doi: 10.1016/j.xcrm.2026.102691

Figure Lengend Snippet: HE4 neutralization exerts therapeutic activity in human cancer models (A–C) PMA-differentiated THP-1 macrophages were stimulated with Fc or hHE4-Fc, and PD-L1 expression was assessed by flow cytometry (A), immunoblotting (B), and RT-qPCR (C); a commercial hHE4-Fc was included as an independent control. (D) Binding of hHE4 to PMA-differentiated THP-1 cells assessed by flow cytometry. (E and F) PMA-differentiated THP-1 cells were pretreated with ruxolitinib, fludarabine, or Stattic, followed by hHE4-Fc stimulation; PD-L1 was quantified by RT-qPCR (E) and flow cytometry (F). (G and H) Anti-hHE4 monoclonal antibodies inhibited hHE4-induced PD-L1 upregulation in PMA-differentiated THP-1 cells, assessed by RT-qPCR (G) and flow cytometry (H). (I) Anti-hHE4 mAb clone #10 blocked hHE4 binding to PMA-differentiated THP-1 cells. (J) Binding of wild-type or epitope-mutant hHE4-Fc to anti-hHE4 mAb clone #10 was quantified by ELISA. (K) Pharmacokinetic analysis of anti-hHE4 mAb clone #10 in C57BL/6 mice following intravenous administration. (L–O) Fresh human LUAD tumor cell suspensions were treated with anti-hHE4 mAb clone #10, followed by flow cytometric analysis of PD-L1 and ELISA measurement of IFN-γ and granzyme B. (P) Recombinant HE4 suppressed IFN-γ production in human LUAD tumor cell suspensions. (Q–T) HE4 blockade enhanced PBMC-mediated antitumor activity in humanized C-NKG mice bearing OVCAR3 or NCI-H358 tumors, shown by treatment scheme, representative tumors, and tumor weights. Schematics (L and Q) were created using BioRender. Statistical analyses were performed using one-way ANOVA (C, E, and G), paired t tests (M–P), or unpaired t tests (S and T). Data in (A–J) are representative of three independent experiments; data in (Q–T) are pooled from two independent experiments.

Article Snippet: Recombinant Fc-tagged mouse IFN-γ protein , MedChemExpress , Cat# HY- P73252.

Techniques: Neutralization, Activity Assay, Expressing, Flow Cytometry, Western Blot, Quantitative RT-PCR, Control, Binding Assay, Bioprocessing, Mutagenesis, Enzyme-linked Immunosorbent Assay, Recombinant