Journal: bioRxiv

Article Title: Integrated kidney and urine proteomics define encrypted antimicrobial peptides as effectors of host defence in human pyelonephritis

doi: 10.64898/2026.04.10.717476

Figure Lengend Snippet: FFPE human kidney sections from PN ( n =3) and control patients ( n =2) were stained with DAPI (nuclei), CD14 (monocytes), CD66b (neutrophils), AQP1 (proximal tubules), NKCC2 (distal tubules), and the following AMPs: AZU1, CTSG, S100A8, and S100A12. (A) Exemplary images of immune and tubular markers as well as AMPs in kidney tissue from patients C2 (Control) and PN3 (Pyelonephritis) (B) Frequency of AMP + CD14 + CD66b - monocytes (red), AMP + CD66b + neutrophils (orange), AMP + AQP1 + stromal cells (turquoise), and AMP + NKCC2 + stromal cells (green) per mm 2 in control (top) and PN (bottom) kidney tissue.

Article Snippet: Antibodies targeted AQP1 (Sigma-Aldrich, HPA019206), NKCC2 (Atlas Antibodies, HPA014967), CD14 (Miltenyi Biotec, 130-110-576), CD66b (Miltenyi Biotec, 130-122-922), AZU1 (Atlas Antibodies, HPA075964), CTSG (Atlas Antibodies, HPA047737), and S100A8 (Sigma-Adrich, HPA024372).

Techniques: Control, Staining

Journal: bioRxiv

Article Title: Integrated kidney and urine proteomics define encrypted antimicrobial peptides as effectors of host defence in human pyelonephritis

doi: 10.64898/2026.04.10.717476

Figure Lengend Snippet: Neutrophils and CD14+ monocytes were isolated from peripheral blood of healthy donors (n = 5) and incubated with 0, 4, or 16 μM Calcitermin for 18 h. Representative dot plots show side scatter characteristics, the proportion of Annexin V+ apoptotic cells, and mean fluorescence intensity (MFI) of markers associated with activation and function, including IL-8, IL-12, CD11b, CD63, CD66b, CXCR4, LAP–TGF-β1, TLR2, and TLR4. Reactive oxygen species (ROS) production was assessed using dihydrorhodamine 123 (DHR 123). Statistical significance is indicated as *p < 0.05 and **p < 0.005.

Article Snippet: Antibodies targeted AQP1 (Sigma-Aldrich, HPA019206), NKCC2 (Atlas Antibodies, HPA014967), CD14 (Miltenyi Biotec, 130-110-576), CD66b (Miltenyi Biotec, 130-122-922), AZU1 (Atlas Antibodies, HPA075964), CTSG (Atlas Antibodies, HPA047737), and S100A8 (Sigma-Adrich, HPA024372).

Techniques: Isolation, Incubation, Fluorescence, Activation Assay

Journal: Nature Communications

Article Title: NCX1 reverse mode promotes calcium-dependent Neutrophil Extracellular Trap formation and lung damage in chronic obstructive pulmonary disease

doi: 10.1038/s41467-026-69636-1

Figure Lengend Snippet: a Representative immunofluorescence images of NCX1 (purple) co-localization with different immune cell-type markers (CD3 + , T cells, CD68 + , macrophages, CD66b + , neutrophils; CD11c + , neutrophils/dendritic cells) in distal airways and bronchioles and adjacent alveolar regions from Mixed CBE patients ( n = 5 patients). Blue, DAPI-stained nuclei. Light blue indicates colocalization of NCX1 with neutrophils. Scale bar = 30 µm. b RT-qPCR analyzes mRNA levels of NCX1 in human lung tissue from Control patients ( n = 20), emphysema patients ( n = 10), and Mixed CBE patients ( n = 20). c Western blot analysis of NCX1 protein levels in lung tissues from Control patients and Mixed CBE patients ( n = 10), normalized to β-actin and displayed relative to controls. Uncropped blots in Source Data. d , e RT-qPCR analyzes mRNA levels of MPO and NE in lung tissues from Control individuals ( n = 20), emphysema patients ( n = 10), and Mixed CBE patients ( n = 20). f–h Western blot analysis of MPO and NE expression in lung tissues from Control patients and Mixed CBE patients ( n = 10), normalized to β-actin and displayed relative to controls. Uncropped blots in Source Data. i , j Spearman’s correlation analyzes the correlation of NCX1 expression with MPO and NE levels. 95% CI: 0.5281 to 0.8724 for ( i ), 0.7029 to 1.007 for ( j ). k Representative immunofluorescence images show expression and localization of CD66b (green) and NCX1 (purple) in lung tissues of Control and Mixed CBE patients. Scale bar = 50 µm. Blue, DAPI-stained nuclei. l–n Flow cytometry identifies neutrophil numbers ( m ) and NCX1 mean fluorescence intensity ( n ) on neutrophils in human BALF ( n = 5 individuals). o Representative immunofluorescence images show expression of NCX1 in neutrophils from human BALF of Control and Mixed CBE patients. Scale bar = 2 µm. Blue, DAPI-stained nuclei. Data point represents one biologically independent replicate with three technical replicates ( b , d , e ), one biologically independent replicate with two technical replicates ( c , g , h ). Data points represent biologically independent replicates ( m , n ). Quantitative data are presented as min to max with all points mean ( b , d , e ), and as Mean ± SD ( c , g , h , m , n ). Two-sided t -test ( g , h , m , n ) and one-way ANOVA with Tukey’s multiple comparison test ( b , d , e ) were used to calculate the p- values. At least 3 times, each experiment was independently repeated with similar results. Source data are provided as a Source Data file. CBE chronic bronchitis and emphysema; MPO myeloperoxidase; NE neutrophil elastase; BALF bronchoalveolar lavage fluid; SCC-A forward scatter-area; FSC-H forward scatter-height; FSC-A side scatter-area; ns no significance.

Article Snippet: The samples were then incubated overnight at 4 °C with primary antibodies: CD11c (Servicebio, GB11059, 1:200), CD68 (Proteintech, 25747-1, 1:200), CD3 (Servicebio, GB11014, 1:100), Ly-6G (Invitrogen, 14-5931-82, 1:50), αSMA (Invitrogen, 50976082, 1:200), CD66b (Novus Biologicals, NB100-77808, 1:100), SFTPC (Proteintech, 10774-1-AP, 1:200), MPO (Proteintech, 22225-1-AP, 1:200), NE (Abclonal, A8953, 1:200), Cit-H3 (Abways, CY6587, 1:250) and NCX1 (Abclonal, A5583, 1:200).

Techniques: Immunofluorescence, Staining, Quantitative RT-PCR, Control, Western Blot, Expressing, Flow Cytometry, Fluorescence, Comparison

Journal: Translational Cancer Research

Article Title: Multi-omics analysis of BTF3L4 as a prognostic and immune biomarker in hepatocellular carcinoma

doi: 10.21037/tcr-2025-aw-2179

Figure Lengend Snippet: Correlation of BTF3L4 mRNA expression with tumor-infiltrating immune cells and immunoregulatory genes in HCC. (A-J) Correlation between BTF3L4 mRNA expression and (A) CTLA-4, (B) CD274 (PD-L1), (C) PDCD1 (PD-1), (D) CD8 + T cells, (E) M1-like macrophages, (F) CD20 + B cells, (G) CD66b + neutrophils, (H) LAMP3 + dendritic cells, (I) CD4 + T cells, and (J) M2-like macrophages by Spearman correlation analysis based on the GEO database. GEO, Gene Expression Omnibus; HCC, hepatocellular carcinoma; mRNA, messenger RNA; PD-1, programmed cell death-1; PD-L1, programmed death-ligand 1.

Article Snippet: The following primary antibodies were used: anti-cytokeratin (CK) antibody (1:1,000, Orb69073, Biorbyt, Cambridge, UK), anti-BTF3L4 antibody (1:250, Orb511711, Biorbyt), anti-CD3e antibody (1:3,000, 85061S, CST, Danvers, MA, USA), anti-CD4 antibody (1:100, ab133616, Abcam, Cambridge, UK), anti-CD8 antibody (1:200, 85336S, CST), anti-CD20 antibody (1:200, ab78237, Abcam), anti-CD66b antibody (1:10,000, arg66287, Arigo, Zhubei, China), anti-CD68 antibody (1:10,000, 76437S, CST), anti-CD86 antibody (1:100, MAB1774, R&D Systems, Minneapolis, MN, USA), anti-CD163 antibody (1:100, 93498S, CST), anti-Lamp3 antibody (1:500, ab111090, Abcam), anti-PD-1 antibody (1:500, 86163, CST), anti-PD-L1 antibody (1:100, 13684S, CST), anti-cytotoxic T-lymphocyte-associated antigen 4 (anti-CTLA-4) (1:2,000, Orb527271, Biorbyt).

Techniques: Expressing, Gene Expression

Journal: Translational Cancer Research

Article Title: Multi-omics analysis of BTF3L4 as a prognostic and immune biomarker in hepatocellular carcinoma

doi: 10.21037/tcr-2025-aw-2179

Figure Lengend Snippet: Representative mIHC images of tumor-infiltrating immune cells and immunoregulatory genes. Representative mIHC images show the fluorescence intensities of the composite of BTF3L4, CTLA-4, CD66b, CD3, CD4, CK and DAPI through seven colors in (A) HCC tissues and (B) adjacent non-tumor tissues. CK, cytokeratin; HCC, hepatocellular carcinoma; mIHC, multiplex immunohistochemistry.

Article Snippet: The following primary antibodies were used: anti-cytokeratin (CK) antibody (1:1,000, Orb69073, Biorbyt, Cambridge, UK), anti-BTF3L4 antibody (1:250, Orb511711, Biorbyt), anti-CD3e antibody (1:3,000, 85061S, CST, Danvers, MA, USA), anti-CD4 antibody (1:100, ab133616, Abcam, Cambridge, UK), anti-CD8 antibody (1:200, 85336S, CST), anti-CD20 antibody (1:200, ab78237, Abcam), anti-CD66b antibody (1:10,000, arg66287, Arigo, Zhubei, China), anti-CD68 antibody (1:10,000, 76437S, CST), anti-CD86 antibody (1:100, MAB1774, R&D Systems, Minneapolis, MN, USA), anti-CD163 antibody (1:100, 93498S, CST), anti-Lamp3 antibody (1:500, ab111090, Abcam), anti-PD-1 antibody (1:500, 86163, CST), anti-PD-L1 antibody (1:100, 13684S, CST), anti-cytotoxic T-lymphocyte-associated antigen 4 (anti-CTLA-4) (1:2,000, Orb527271, Biorbyt).

Techniques: Fluorescence, Multiplex Assay, Immunohistochemistry

Journal: Translational Cancer Research

Article Title: Multi-omics analysis of BTF3L4 as a prognostic and immune biomarker in hepatocellular carcinoma

doi: 10.21037/tcr-2025-aw-2179

Figure Lengend Snippet: Correlation of BTF3L4 protein expression with tumor-infiltrating immune cells (TIICs) and immunoregulatory genes in HCC. (A) Heatmap of the association between BTF3L4 protein levels and TIICs as well as immunoregulatory genes. (B-K) Correlation between BTF3L4 protein levels and (B) CD66b + neutrophils, (C) CD4 + T cells (CD3 + CD4 + ), (D) CTLA-4, (E) CD20 + B cells, (F) M1-like macrophages (CD86 + CD68 + ), (G) M2-like macrophages (CD163 + CD68 + ), (H) CD8 + T cells (CD3 + CD8 + ), (I) LAMP3 + dendritic cells, (J) PD-1, and (K) PD-L1 by Spearman correlation analysis. HCC, hepatocellular carcinoma; PD-1, programmed cell death-1; PD-L1, programmed death-ligand 1; TIICs, tumor-infiltrating immune cells.

Article Snippet: The following primary antibodies were used: anti-cytokeratin (CK) antibody (1:1,000, Orb69073, Biorbyt, Cambridge, UK), anti-BTF3L4 antibody (1:250, Orb511711, Biorbyt), anti-CD3e antibody (1:3,000, 85061S, CST, Danvers, MA, USA), anti-CD4 antibody (1:100, ab133616, Abcam, Cambridge, UK), anti-CD8 antibody (1:200, 85336S, CST), anti-CD20 antibody (1:200, ab78237, Abcam), anti-CD66b antibody (1:10,000, arg66287, Arigo, Zhubei, China), anti-CD68 antibody (1:10,000, 76437S, CST), anti-CD86 antibody (1:100, MAB1774, R&D Systems, Minneapolis, MN, USA), anti-CD163 antibody (1:100, 93498S, CST), anti-Lamp3 antibody (1:500, ab111090, Abcam), anti-PD-1 antibody (1:500, 86163, CST), anti-PD-L1 antibody (1:100, 13684S, CST), anti-cytotoxic T-lymphocyte-associated antigen 4 (anti-CTLA-4) (1:2,000, Orb527271, Biorbyt).

Techniques: Expressing