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

Journal: Cell Communication and Signaling : CCS

Article Title: Interferon-alpha selectively signals mitochondrial pore opening to allow mitochondrial RNA release in systemic lupus erythematosus: pathophysiological implications

doi: 10.1186/s12964-026-02910-3

Figure Lengend Snippet: Induction of mtRNA leakage by IFN-α-100 and ICs treatment. BMDMs (2 × 10 6 ) were stimulated with IFN-α (100 U/ml) or 10 µg/ml ICs for 24 h, and the levels of cytosolic mtRNA were determined by measuring both the heavy chain and the light chain of several mtRNA genes as indicated by RT-qPCR ( A and C ) and confocal microscopy ( B and D ). The heavy and light chains of some mtRNA genes in the supernatant were determined in BMDMs treated with IFN-α (100 U/ml) or 10 µg/ml ICs for different periods of time ( E and F ). The levels of cytosolic mtRNA in BMDMs treated with anti-chicken egg albumin Abs, albumin (ovalbumin) or a combination of anti-albumin Abs and albumin (ICs) were determined ( G ). Each data point represents one mouse, and the values are fold changes relative to the mean value of the unstimulated controls determined by RT-qPCR ( A , C , E , F , and G ). Representative results from at least 3 independent experiments are shown ( B and D ). Statistical analysis was performed with unpaired Student’s t test to compare the means between two groups ( A , C , E , and F ) and two-way ANOVA with the Holm-Šídák’s multiple comparisons test to compare differences among different treatments ( G ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ND, NADH dehydrogenase; CYTB, cytochrome b

Article Snippet: Recombinant mouse IFN-α (12100–1) was purchased from PBL Assay Science (Piscataway, NJ, USA).

Techniques: Quantitative RT-PCR, Confocal Microscopy

Journal: Cell Communication and Signaling : CCS

Article Title: Interferon-alpha selectively signals mitochondrial pore opening to allow mitochondrial RNA release in systemic lupus erythematosus: pathophysiological implications

doi: 10.1186/s12964-026-02910-3

Figure Lengend Snippet: IFN-α-100 and ICs treatment selectively induces mitochondrial pore opening. BMDMs (2 × 10 6 ) were stimulated with IFN-α (100 U/ml) ( A ) or 10 µg/ml ICs ( B ) for 24 h, and mPTP opening was measured by calcein AM staining, and the results were analyzed by flow cytometry as described in the Materials and Methods. Images of calcein AM staining were analyzed by confocal microscopy ( C ). The protein levels of ANT1 and ANT2 were determined ( D ). The oligomerization of VDAC1 ( E and F ) and BAX ( G and H ) with or without EGS, a cross-linking reagent used to stabilize oligomers in electrophoresis, was evaluated, and the results of the statistical analysis are presented individually ( E to H ). As a positive control for BAX oligomerization, the treatment with LPS (200 ng/ml) for 4 h and then ATP (4 mM) for 1 h ( G ) or TNF-α (20 ng/ml) plus cycloheximide (CHX; 1 µg/ml) for 3 h ( H ) was given. BMDMs (2 × 10 6 ) were pre-treated with BAI-1 (2 µM) for 2 h and then stimulated with IFN-α (100 U/ml) or 10 µg/ml ICs for 24 h. As positive controls, BMDMs were pre-treated with BAI-1 (2 µM) for 2 h and then stimulated with staurosporine (STS; 2 µM) or TNF-α (20 ng/ml) plus cycloheximide (CHX; 30 µg/ml) for 3 h. MtRNA release was measured by RT-qPCR ( I ). Each data point represents one mouse, and the values for the flow cytometry results are presented as the geomeans (MFIs) ( A and B ). For Western blotting, the ratio of protein levels was calculated by comparing the intensity of a protein of interest to that of a housekeeping protein or monomer of protein as indicated ( D to H ). The samples were derived from the same experiment, and both the gels and the blots were processed in parallel. In ( I ), the values are fold changes relative to the mean value of the controls determined in RT-qPCR. Statistical analysis was performed with an unpaired Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ANT, adenine nucleotide translocase; Tri, trimer; Di, dimer; PDTC, pyrrolidine dithiocarbamate ammonium

Article Snippet: Recombinant mouse IFN-α (12100–1) was purchased from PBL Assay Science (Piscataway, NJ, USA).

Techniques: Staining, Flow Cytometry, Confocal Microscopy, Electrophoresis, Positive Control, Quantitative RT-PCR, Western Blot, Derivative Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Interferon-alpha selectively signals mitochondrial pore opening to allow mitochondrial RNA release in systemic lupus erythematosus: pathophysiological implications

doi: 10.1186/s12964-026-02910-3

Figure Lengend Snippet: Inhibition of mitophagy does not block IFN-α-100 and ICs-induced mtRNA release. BMDMs (2 × 10 6 ) were stimulated with IFN-α (100 U/ml) or 10 µg/ml ICs for 24 h, and the intensity of mitophagy was determined by flow cytometry as described in the Materials and Methods section, and the statistics are presented ( A and B ). C The results of the confocal microscopic imaging studies. D and E show the inhibitory effects of Mdivi-1 (50 µM), a mitophagy inhibitor, on IFN-α-100 and ICs-induced mitophagy by flow cytometry. Mdivi-1 did not affect cytosolic mtRNA release, as determined by measuring ND1 and Co-2 mRNA expression in the cytosol ( F and G ). Each data point represents one mouse, and the values for the flow cytometry results are presented as the geomeans (MFIs) ( A , B , D , and E ). The values are fold changes relative to the mean value of the controls in RT‒qPCR ( F and G ). Representative results from at least 3 independent experiments are shown ( C ). Statistical analysis was performed with unpaired Student’s t test to compare the means between two groups ( A and B ). Two-way ANOVA with Holm-Šídák’s multiple comparisons was used to compare differences among different treatments ( D to G ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. The arrows show the colocalization of mitophagy dye and lysosomal marker

Article Snippet: Recombinant mouse IFN-α (12100–1) was purchased from PBL Assay Science (Piscataway, NJ, USA).

Techniques: Inhibition, Blocking Assay, Flow Cytometry, Imaging, Expressing, Marker

Journal: Cell Communication and Signaling : CCS

Article Title: Interferon-alpha selectively signals mitochondrial pore opening to allow mitochondrial RNA release in systemic lupus erythematosus: pathophysiological implications

doi: 10.1186/s12964-026-02910-3

Figure Lengend Snippet: Inhibition of mPTP opening and VDAC1 oligomerization attenuates IFN-α-100 and ICs-induced mtRNA release and downstream signaling. BMDMs (2 × 10 6 ) were pretreated with cyclosporin A (5 µM) or VBIT-12 (80 µM) for 2 h and then stimulated with IFN-α (100 U/ml) or 10 µg/ml ICs for 24 h. The intensity of mPTP opening ( A and D ), mtRNA release ( B , E , H , and K ), mRNA levels of several inflammatory genes ( C , F , I , and L ), and VDAC1 oligomerization ( G and J ) were measured as described in Materials and Methods. Each data point represents one mouse, and the values for the flow cytometry results are presented as the geomeans (MFIs) ( A and D ). The values are fold changes relative to the mean value of the controls as determined by RT-qPCR ( B , C , E , F , H , I , K , and L ). Representative results from at least 3 independent experiments are shown ( G and J ). Statistical analysis was performed with unpaired Student’s t test to compare the means between two groups ( C , F , I , and L ). Two-way ANOVA with Holm-Šídák’s multiple comparisons was used to compare differences among different treatments ( A , B , D , E , H , and K ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. RIG1, retinoic acid-inducible gene I; ISG15, interferon-stimulated gene 15; IL, interleukin; MDA5, melanoma differentiation-associated protein 5; OAS, 2′-5′-oligoadenylate synthetase; Tri, trimer; Di, dimer

Article Snippet: Recombinant mouse IFN-α (12100–1) was purchased from PBL Assay Science (Piscataway, NJ, USA).

Techniques: Inhibition, Flow Cytometry, Quantitative RT-PCR

Journal: Cell Communication and Signaling : CCS

Article Title: Interferon-alpha selectively signals mitochondrial pore opening to allow mitochondrial RNA release in systemic lupus erythematosus: pathophysiological implications

doi: 10.1186/s12964-026-02910-3

Figure Lengend Snippet: Blockade of mitochondrial calcium overload inhibits IFN-α-100 and ICs-induced mtRNA release. BMDMs (2 × 10 6 ) were pretreated with RuR (30 µM) for 2 h and then stimulated with IFN-α (100 U/ml) or 10 µg/ml ICs for 24 h. The mitochondrial Ca 2+ concentration ([Ca 2+ ]m) was measured by Rhod-2 AM staining and analyzed by flow cytometry ( A and B ). ( C ) shows the results of confocal microscopy. mPTP opening was determined with calcein AM staining and analyzed by flow cytometry ( D and E ). The levels of cytosolic mtRNA, ND1 and Co-2, ( F and H ) or mRNA of inflammatory signaling molecules ( G and I ) were measured by RT-qPCR. Each data point represents one mouse, and the values for the flow cytometry results are presented as the geomeans (MFIs) ( A , B , D , and E ). The values are fold changes relative to the mean value of the controls according to the results from RT-qPCR ( F , G , H , and I ). Representative results from at least 3 independent experiments are shown ( C ). Statistical analysis was performed with unpaired Student’s t test to compare the means between two groups ( G and I ). Two-way ANOVA with Holm-Šídák’s multiple comparisons was used to compare differences among different treatments ( A , B , D , E , F , and H ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. RIG1, retinoic acid-inducible gene I; ISG15, interferon-stimulated gene 15; IL, interleukin; MDA5, melanoma differentiation-associated protein 5; OAS, 2′-5′-oligoadenylate synthetase, TNF-α, tumor necrosis factor-α

Article Snippet: Recombinant mouse IFN-α (12100–1) was purchased from PBL Assay Science (Piscataway, NJ, USA).

Techniques: Concentration Assay, Staining, Flow Cytometry, Confocal Microscopy, Quantitative RT-PCR

Journal: Cell Communication and Signaling : CCS

Article Title: Interferon-alpha selectively signals mitochondrial pore opening to allow mitochondrial RNA release in systemic lupus erythematosus: pathophysiological implications

doi: 10.1186/s12964-026-02910-3

Figure Lengend Snippet: Endoplasmic reticulum calcium release regulates IFN-α-100 and ICs-induced mtRNA release. BMDMs (2 × 10 ) were pretreated with XeC (5 µM) for 2 h and then stimulated with IFN-α (100 U/ml) or 10 µg/ml ICs for 24 h. The signal and mitochondrial Ca 2+ concentrations ([Ca 2+ ]m) ( A , B , and C ), mPTP opening ( D and E ), cytosolic mtRNA levels ( F and H ), and mRNA levels of inflammatory signaling molecules ( G and I ) were measured as described in Fig. . Each data point represents one mouse, and the values for the flow cytometry results are presented as the geomeans (MFIs) ( A , B , D , and E ). The values are fold changes relative to the mean value of the controls ( F , G , H , and I ). Representative results from at least 3 independent experiments are shown ( C ). Statistical analysis was performed with unpaired Student’s t test to compare the means between two groups ( G and I ). Two-way ANOVA with Holm-Šídák’s multiple comparisons was used to compare differences among different treatments ( A , B , D , E , F , and H ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. RIG1, retinoic acid-inducible gene I; ISG15, interferon-stimulated gene 15; IL, interleukin; MDA5, melanoma differentiation-associated protein 5; OAS, 2′-5′-oligoadenylate synthetase, TNF-α, tumor necrosis factor-α

Article Snippet: Recombinant mouse IFN-α (12100–1) was purchased from PBL Assay Science (Piscataway, NJ, USA).

Techniques: Flow Cytometry

Journal: Cell Communication and Signaling : CCS

Article Title: Interferon-alpha selectively signals mitochondrial pore opening to allow mitochondrial RNA release in systemic lupus erythematosus: pathophysiological implications

doi: 10.1186/s12964-026-02910-3

Figure Lengend Snippet: Effects of IFN-α-100 or ICs treatment on mitochondrial membrane potential, apoptosis and cell viability. BMDMs (2 × 10 6 ) were treated with IFN-α (100 U/ml) or 10 µg/ml ICs for 24 h. To measure changes in mitochondrial membrane potential, JC-1 green/red staining was performed, after which the cells were analyzed by flow cytometry ( A ). Several approaches to measure cell viability, apoptosis and cell death as described in Materials and Methods were performed. These include the measurements of SYTOX staining ( B ), LDH release ( C ), CCK-8 assays ( D ), annexin-V staining ( E ), and ATP levels ( F ). BMDMs (2 × 10 6 ) were pre-treated with Z-VAD-FMK (20 µM) for 2 h and then stimulated with IFN-α (100 U/ml) or 10 µg/ml ICs for 24 h. As controls, BMDMs were pre-treated with Z-VAD-FMK (20 µM) for 2 h and then stimulated with TNF-α (20 ng/ml) plus cycloheximide (CHX; 30 µg/ml) for 3 h. The cleavaged products from PARP and caspase 3 were detected by Western blotting ( G ). Cytosolic mtRNA release was measured by RT-qPCR ( H ). Each data point represents one mouse, and the values are fold changes relative to the mean value of the controls ( A and F ). For measuring SYTOX staining and annexin V staining, the values are the percentages of cells with SYTOX Green-positive nuclei (pseudocolored red) or annexin V-positive relative to the total cell population ( B and E ). The values of LDH release are the percentage relative to the mean value of the positive controls ( C ). The values of the CCK-8 and ATP level are the readouts from the absorbance microplate reader and luminescence reader ( D and F ). Representative results from at least 3 independent experiments are shown ( B and G ). One-way ANOVA ( B , D and F ) or two-way ANOVA ( A , C , E and H ) with Holm-Šídák’s multiple comparisons was used to compare differences among different treatments. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Recombinant mouse IFN-α (12100–1) was purchased from PBL Assay Science (Piscataway, NJ, USA).

Techniques: Membrane, Staining, Flow Cytometry, CCK-8 Assay, Western Blot, Quantitative RT-PCR, Microplate Reader Absorbance Measurement

Journal: Cell Communication and Signaling : CCS

Article Title: Interferon-alpha selectively signals mitochondrial pore opening to allow mitochondrial RNA release in systemic lupus erythematosus: pathophysiological implications

doi: 10.1186/s12964-026-02910-3

Figure Lengend Snippet: The sequential events involved in the regulation of mtRNA release into the cytosol by IFN-α-100 and IC treatment. Stimulation of IFN-α-100 and ICs resulted in the release of calcium from the endoplasmic reticulum that entered the mitochondria, induced mitochondrial calcium overload, and triggered the opening of mitochondrial pores, including the oligomerization of voltage dependent anion channel (VDAC) and mPTP. The activation of adenine nucleotide translocase (ANT) also participated in the opening of mitochondrial pores in the inner mitochondrial membrane. Although ANT is not an essential component of the mPTP, ANT can regulate mPTP opening. The opening of the mPTP in the inner mitochondrial membrane and the oligomerization of VDAC1 resulted in the release of mtRNA into the cytosol, which activated its sensors RIG-I and MDA5 as well as several downstream inflammatory signaling pathways to induce innate immunity

Article Snippet: Recombinant mouse IFN-α (12100–1) was purchased from PBL Assay Science (Piscataway, NJ, USA).

Techniques: Activation Assay, Membrane, Protein-Protein interactions

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

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