Journal: Science Advances

Article Title: A robust adhesive microneedle for oral infections therapy via synergistic antibacterial and neutrophil-macrophage axis immunomodulation

doi: 10.1126/sciadv.aee4401

Figure Lengend Snippet: ( A ) Fluorescence images of S. aureus stained with C11 BODIPY 581/591 for LPO detection. ( B ) Chemical structures of POPG and OOPG. ( C ) Side and top views of bacterial membrane thickness. ( D ) An MD simulation snapshot depicting the transmembrane diffusion behavior of CO. ( E ) MSD analysis used to calculate the diffusion coefficient of CO. ( F ) TEM images of the bacterial microstructure. ( G ) Volcano plot of DEGs. ( H ) Heatmap of the differentially expressed genes. ( I ) GO functional enrichment analysis of DEGs. ( J ) GSEA of the transcriptomic data. ( K ) Activity analysis of ETC complexes I to IV. Data are presented as means ± SD ( n = 3 to 4 biologically independent samples).

Article Snippet: C11 BODIPY 581/591 was purchased from MedChemExpress (Shanghai, China).

Techniques: Fluorescence, Staining, Membrane, Diffusion-based Assay, Functional Assay, Activity Assay

Journal: Oncology Letters

Article Title: Lung squamous cell carcinoma downregulates haptoglobin expression to inhibit M2 macrophage ferroptosis via the hemoglobin-dependent CD163/HO-1 pathway

doi: 10.3892/ol.2026.15558

Figure Lengend Snippet: Overexpression of HP in LUSC cells does not induce ferroptosis in LUSC cells. (A and B) NCI-H226 and NCI-H520 cells were infected with LV- HP or LV-Ctrl. After 72 h of infection, cells and culture supernatants were collected for analysis of HP (A) mRNA and (B) protein levels by RT-PCR and ELISA, respectively. (C) NCI-H226 and NCI-H520 cells infected with LV- HP or LV-Ctrl were cultured with or without Hb (30 µg/ml) for 24 h. Culture supernatants were collected and LDH release was measured. (D) Cells were analyzed via flow cytometry to determine the percentage of 7-AAD-positive cells in NCI-H226 and NCI-H520 cells. (E and F) The mean fluorescence intensity of (E) FerroOrange (Fe 2+ ) and (F) C11-BODIPY (lipid peroxide) was measured using flow cytometry. Data are presented as the mean ± SD from five independent experiments and were analyzed using an unpaired t-test or a two-way ANOVA followed by a Tukey's post-hoc multiple comparison analysis. ****P<0.0001. ns, not significant; HP, haptoglobin; Hb, hemoglobin; LDH, lactate dehydrogenase; 7-AAD, 7-aminoactinomycin D.

Article Snippet: Phorbol 12-Myristate 13-Acetate (PMA; cat. no. HY-18739), FerroOrange dye (cat. no. HY-D1913), BODIPY 581/591 C11 dye (cat. no. HY-D1301), tin protoporphyrin IX dichloride (SnPPIX, cat. no. HY-101194) and deferoxamine (DFO; cat. no. HY-B1625) were purchased from MedChemExpress.

Techniques: Over Expression, Infection, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Cell Culture, Flow Cytometry, Fluorescence, Comparison

Journal: Oncology Letters

Article Title: Lung squamous cell carcinoma downregulates haptoglobin expression to inhibit M2 macrophage ferroptosis via the hemoglobin-dependent CD163/HO-1 pathway

doi: 10.3892/ol.2026.15558

Figure Lengend Snippet: CM from LUSC cells overexpressing HP leads to ferroptosis in M2 macrophages. (A) THP-1 cells were differentiated into macrophages by treating them with 150 nM PMA for 24 h, followed by culture in fresh RPMI-1640 for 24 h. The macrophages were polarized into M2 macrophages by incubating with IL-4 (20 ng/ml) and IL-13 (20 ng/ml) for 24 h. Flow cytometry was then performed to identify CD163 + CD206 + M2 macrophages. (B) M2 macrophages were cultured with the CM from NCI-H226 and NCI-H520 cells infected with LV- HP or LV-Ctrl in the presence or absence of Hb for 24 h. LDH release was analyzed by ELISA. Treated M2 macrophages were treated as in (B) and flow cytometry was used to detect the proportion of (C) 7-AAD positive cells, as well as the mean fluorescence intensity of (D) FerroOrange and (E) C11-BODIPY. (F) Treated M2 macrophages were observed using transmission electron microscopy. Scale bar, 5 µm. Data are presented as the mean ± SD from five independent experiments and were analyzed using an unpaired t-test or a two-way ANOVA followed by a Tukey's post-hoc multiple comparison analysis. ****P<0.0001. ns, not significant; CM, conditioned media; LUSC, lung squamous cell carcinoma; LDH, lactate dehydrogenase; 7-AAD, 7-aminoactinomycin D; HP, haptoglobin.

Article Snippet: Phorbol 12-Myristate 13-Acetate (PMA; cat. no. HY-18739), FerroOrange dye (cat. no. HY-D1913), BODIPY 581/591 C11 dye (cat. no. HY-D1301), tin protoporphyrin IX dichloride (SnPPIX, cat. no. HY-101194) and deferoxamine (DFO; cat. no. HY-B1625) were purchased from MedChemExpress.

Techniques: Flow Cytometry, Cell Culture, Infection, Enzyme-linked Immunosorbent Assay, Fluorescence, Transmission Assay, Electron Microscopy, Comparison

Journal: Oncology Letters

Article Title: Lung squamous cell carcinoma downregulates haptoglobin expression to inhibit M2 macrophage ferroptosis via the hemoglobin-dependent CD163/HO-1 pathway

doi: 10.3892/ol.2026.15558

Figure Lengend Snippet: HP promotes ferroptosis in M2 macrophage by inducing intracellular Fe 2+ generation via the CD163/HO-1 signaling pathway. (A and B) M2 macrophages were cultured with CM from NCI-H226 and NCI-H520 cells infected with LV- HP or LV-Ctrl, in the presence of Hb and an anti-CD163 antibody. The heme levels in M2 macrophages were measured using (A) spectrophotometry and (B) HO-1 protein expression levels were examined using flow cytometry. (C) M2 macrophages were co-cultured with CM containing Hb, with or without the addition of an anti-CD163 antibody, the HO-1 inhibitor SnPPIX, or the Fe 2+ chelator DFO for 24 h. (D) Following treatment with the anti-CD163 antibody, the percentage of 7-AAD + cells, and the mean fluorescence intensity of FerroOrange and (E) C11-BODIPY were analyzed by flow cytometry. After SnPPIX treatment, (F) 7-AAD + cells, (G) FerroOrange intensity (H) and C11-BODIPY intensity were assessed. Following DFO treatment, (I) 7-AAD + cells, (J) FerroOrange intensity, and (K) C11-BODIPY intensity were measured. Data are presented as the mean ± SD from five independent experiments and were analyzed using an unpaired t-test or a two-way ANOVA followed by a Tukey's post-hoc multiple comparison analysis. ****P<0.0001. HP, haptoglobin; CM, conditioned media; Hb, hemoglobin; 7-AAD, 7-aminoactinomycin D; HO-1, heme oxygenase-1; DFO, deferoxamine.

Article Snippet: Phorbol 12-Myristate 13-Acetate (PMA; cat. no. HY-18739), FerroOrange dye (cat. no. HY-D1913), BODIPY 581/591 C11 dye (cat. no. HY-D1301), tin protoporphyrin IX dichloride (SnPPIX, cat. no. HY-101194) and deferoxamine (DFO; cat. no. HY-B1625) were purchased from MedChemExpress.

Techniques: Cell Culture, Infection, Spectrophotometry, Expressing, Flow Cytometry, Fluorescence, Comparison