antibodies against inos  (Proteintech)

Journal: Bioactive Materials

Article Title: Integrated apoptotic extracellular vesicle-recruitment peptide coating reprograms the diabetic bone microenvironment and orchestrates enhanced implant osseointegration

doi: 10.1016/j.bioactmat.2026.05.059

Figure Lengend Snippet: In vivo immunomodulatory effects of different modified surfaces in a DM model. (A) Schematic representation of the animal modeling and experimental treatment workflow. (B, C) hematoxylin and eosin staining of the peri-implant tissues in the femurs of DM rats 1 week after implantation, accompanied by quantitative analysis of the fibrous capsule thickness (scale bar = 100 μm, n = 5). (D–G) Immunofluorescence staining evaluating the polarization state of macrophages surrounding the implants (green: macrophage marker cluster of differentiation (CD) 68; red: M1 marker CD86 and M2 marker CD206; blue: nuclei), along with corresponding quantitative analysis of the fluorescence signals (scale bar = 100 μm, n = 5). (H–K) Immunohistochemical staining assessing the expression of the pro-inflammatory marker tumor necrosis factor-α and the anti-inflammatory marker interleukin-10 in the peri-implant area, with quantitative results of the positive staining areas (scale bar = 100 μm, n = 5). Data are expressed as the mean ± standard deviation, with statistical analysis performed using one-way ANOVA and Tukey's post-hoc test. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicate statistical significance.

Article Snippet: For IHC analysis, sections underwent heat-induced antigen retrieval and blocking prior to incubation with antibodies against TNF-α (GB11188, Servicebio, China), IL-10 (GB11534, Servicebio, China), and VEGF (GB15165, Servicebio, China) to identify differences in local inflammatory and angiogenic profiles.

Techniques: In Vivo, Modification, Staining, Immunofluorescence, Marker, Fluorescence, Immunohistochemical staining, Expressing, Standard Deviation

Journal: Advanced Science

Article Title: Redirecting Monocyte Differentiation With Engineered Extracellular Vesicles for Glioma Immunotherapy

doi: 10.1002/advs.76910

Figure Lengend Snippet: C‐EVs‐mediated enhanced BBB permeability, excellent targeting ability, and brain tumor accumulation. (A) Schematic illustration of in vitro BBB model. (B) Transport ratio of liposome and EVs traverses the BBB model after different time periods ( n = 3). (C) Confocal images and relative fluorescence intensity of GL261‐IL13Rα2 cells in the lower chamber after treated with DiD‐labeled liposome or EVs for 24 h ( n = 5). Scale bar, 10 µm. (D) Flow cytometry analysis of fluorescence intensity of GL261‐IL13Rα2 in the lower chamber after incubated with EVs or EVs with anti‐CCR2 ( n = 3). (E) Schematic illustration of the 3D tumor spheroids and penetration of DiD‐labeled liposome or EVs into GL261‐IL13Rα2 tumor spheroids after 4 h incubation. Scale bar, 50 µm. (F) In vivo and ex vivo bioluminescence and fluorescence imaging of GL261‐IL13Rα2 tumor‐bearing mice and brain at 24 h after tail vein injection of DiR‐labeled EVs. Immunofluorescence staining of tumor‐bearing brain, dotted lines demarcate the tumor boundary (T), with adjacent normal brain tissue (N) shown for anatomical reference. Scale bars: 50 µm. (G and H) Representative fluorescence images (G) and their quantitative analysis (H) of GL261‐IL13Rα2‐bearing mice after i.v . injection with free DiR, DiR‐labeled liposome, or EVs at different time points. (I) Ex vivo images of the GL261‐IL13Rα2 bearing brain and their quantification of the fluorescence signal in the brain ( n = 3). (J) Immunofluorescence staining and the corresponding line profiles of the tumor‐bearing brain after tail vein injection of free DiR, DiR‐labeled liposome, or EVs. DAPI (blue) stained nuclei, and CD31 (green) labeled blood vessels. Scale bar, 50 µm. (K) C‐EVs bound to the membrane of GL261‐IL13Rα2 cells. Scale bar, 10 µm. (L) Degree of cellular uptake of EVs and C‐EVs in GL261 and GL261‐IL13Rα2 quantified by flow cytometry. (M) CLSM images of GL261‐IL13Rα2 cells incubated with EVs and C‐EVs at 4 h. Scale bar, 10 µm. (N) In vivo and ex vivo fluorescence imaging, and their quantification of the fluorescence signal of GL261‐IL13Rα2 tumor‐bearing mice and brain at 24 h after tail vein injection of DiR‐labeled EVs or C‐EVs. (O) Immunofluorescence staining of tumor‐bearing brain after i.v . injection with DiR‐labeled EVs or C‐EVs. Scale bars: 100 µm. Statistical analysis was performed by unpaired two‐tailed t ‐test (C,D and N) or one‐way ANOVA with Tukey's multiple comparisons tests (I). The experimental data were presented as mean ± S.E.M. ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Neutralizing antibodies against CCR2 (MedChemExpress, USA) were used in antibody‐blocking experiments.

Techniques: Permeability, In Vitro, Fluorescence, Labeling, Flow Cytometry, Incubation, In Vivo, Ex Vivo, Imaging, Injection, Immunofluorescence, Staining, Membrane, Two Tailed Test

Journal: Bioactive Materials

Article Title: Immunomodulatory effects of biodegradable Mg–Cu–Zn alloy in esophageal cancer

doi: 10.1016/j.bioactmat.2026.02.046

Figure Lengend Snippet: Distribution of CD163 + M2 TAMs in AKR-derived allograft tumor tissues from immunocompetent C57BL/6 mice. (a, b) Representative IHC staining images showing CD163 + M2 TAMs in the (a) peritumoral stroma and (b) tumor islets. Lower panels display higher-magnification views of the regions outlined by red dashed boxes. (c, d) Quantification of CD163 + cells in the (c) peritumoral stroma and (d) tumor islets. (e) Comparison of CD163 + cell density between the peritumoral stroma and tumor islets. (f) Total number of CD163 + cells in allograft tumors (peritumoral stroma and tumor islets combined). (g, h) Comparison of the density between iNOS + cells and CD163 + cells in the (g) peritumoral stroma and (h) tumor islets. (i, j) Quantification of iNOS + /CD163 + ratio in the (i) peritumoral stroma and (j) tumor islets. p < 0.05 (∗), p < 0.01 (∗∗), p < 0.001 (∗∗∗). A field of view is ∼0.086 mm 2 in (c−j).

Article Snippet: Tissue sections were then incubated with primary antibodies against iNOS (22226-1-AP, ProteinTech, China), CD163 (A26411PM, Abclone, China), CD8 (SP16, Maixin, China), CD4 (SP35, Maixin, China) or Ki-67 (12202S, Cell Signaling Technology) for 12 h at 4 °C, followed by secondary antibodies (Beyotime Biotechnology, Nantong, China).

Techniques: Derivative Assay, Immunohistochemistry, Comparison

Journal: Journal of Bone Oncology

Article Title: Lapatinib induces ferroptosis in osteosarcoma via the SLC1A5-GPX4 axis

doi: 10.1016/j.jbo.2026.100771

Figure Lengend Snippet: The SLC1A5-GPX4 axis mediates lapatinib-induced ferroptosis. (A-B) Western blot analysis of SLC1A5 protein expression in OS cells treated with Lap or Lap+DFO (100 μM). (C) Evaluation of the interaction between Lap and SLC1A5 using CETSA. (D) DARTS assay confirming the interaction between Lap and SLC1A5. (E) Viability assessment by calcein-AM (live, green)/PI (dead, red) staining in cells overexpressing SLC1A5. (F) Detection of intracellular Fe 2+ by FerroOrange probe. (G-H) Measurements of Gln and GSH levels in OS cells under indicated conditions. (I) Western blot analysis of SLC1A5 and GPX4 expression following Lap treatment in SLC1A5-overexpressing cells. (J) Western blot analysis of SLC1A5 protein expression in OS cells transfected with control siRNA or siRNA SLC1A5, showing knockdown efficiency. (K) GPX4 protein expression after lapatinib treatment for 24 h in control and SLC1A5 knockdown cells. (L) Cell viability determined by CCK-8 assay. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The membranes were blocked with 5% non-fat milk at room temperature for 1 h, then incubated overnight at 4 °C with primary antibodies against GPX4 (1:5000, Abmart, T56959 ), SLC1A5 (1:2500, Selleck, F0619), and β-actin (1:10000, Servicebio, GB11001).

Techniques: Western Blot, Expressing, Staining, Transfection, Control, Knockdown, CCK-8 Assay

Journal: Journal of Bone Oncology

Article Title: Lapatinib induces ferroptosis in osteosarcoma via the SLC1A5-GPX4 axis

doi: 10.1016/j.jbo.2026.100771

Figure Lengend Snippet: Lapatinib induces ferroptosis by suppressing GPX4. (A-B) Western blot analysis of GPX4 protein expression in OS cells following Lap treatment. (C—D) Western blot analysis of GPX4 expression in OS cells treated with Lap and DFO (100 μM). (E-F) Assessment of cell death by propidium iodide (PI) staining following GPX4 overexpression. (G-H) Measurement of intracellular ROS levels using DCFH-DA probe. (I-J) Flow cytometric analysis of lipid ROS using C11-BODIPY probe.

Article Snippet: The membranes were blocked with 5% non-fat milk at room temperature for 1 h, then incubated overnight at 4 °C with primary antibodies against GPX4 (1:5000, Abmart, T56959 ), SLC1A5 (1:2500, Selleck, F0619), and β-actin (1:10000, Servicebio, GB11001).

Techniques: Western Blot, Expressing, Staining, Over Expression

Journal: Journal of Bone Oncology

Article Title: Lapatinib induces ferroptosis in osteosarcoma via the SLC1A5-GPX4 axis

doi: 10.1016/j.jbo.2026.100771

Figure Lengend Snippet: Lapatinib suppresses tumor growth by activating ferroptosis in vivo. (A) Experimental timeline for in vivo administration of Lap (25 mg/kg, i.p., daily) and DFO (10 mg/kg, i.p., daily). (B—C) Representative photographs of resected xenograft tumors from each group ( n = 4) (Scale bar: 1 cm). (D-E) Tumor weight and volume at the study endpoint. (F-G) Curves depicting body weight changes and tumor growth over time. (H) Hematoxylin and eosin (H&E) staining and immunohistochemical (IHC) analysis of tumor sections (scale bar: 50 μm). (I) Western blot analysis of SLC1A5 and GPX4 protein levels in tumor tissues. (J-K) Measurement of GSH and MDA levels in mouse plasma.

Article Snippet: The membranes were blocked with 5% non-fat milk at room temperature for 1 h, then incubated overnight at 4 °C with primary antibodies against GPX4 (1:5000, Abmart, T56959 ), SLC1A5 (1:2500, Selleck, F0619), and β-actin (1:10000, Servicebio, GB11001).

Techniques: In Vivo, Staining, Immunohistochemical staining, Western Blot, Clinical Proteomics

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 depletion in macrophages restrains LUAD growth and metastasis in vivo. (A) Schematic of orthotopic lung implantation and experimental metastasis models using LLC cells mixed with BMDMs expressing shNC or sh-circSMAD4. (B) Representative images of orthotopic lung tumors. (C) Tumor weight of orthotopic implants. (D) Overall survival of mice bearing orthotopic tumors. (E) Immunofluorescence showing F4/80 and circSMAD4 signals in tumor tissues. Scale bar, 50 μm. (F, G) Representative Ki-67 IHC staining and quantification in orthotopic tumors. Scale bar, 50 μm. (H) Representative bioluminescence images of lung tumor burden in the metastasis model. (I) Tumor weight in the metastasis model. (J) Overall survival of mice in the metastasis model. (K–M) Representative IHC staining and quantification of E-cadherin and vimentin in tumors. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: In Vivo, Expressing, Immunofluorescence, Immunohistochemistry