Journal: Kidney International Reports

Article Title: Flow-Cytometric Quantification of Urine Kidney Epithelial Cells Specifically Reflects Tubular Damage in Acute Kidney Diseases

doi: 10.1016/j.ekir.2025.01.037

Figure Lengend Snippet: The amount of urinary TECs can be measured using flow cytometry. (a) Gating strategy for selecting cytokeratin positive single cells. Staining of CD10, CD13, CD326, and (b) CD227 compared with isotype-controls in postmortem kidney tissue, (c) postmortem ureteral tissue, and (d) urine. Note CD326 but not CD227 positivity in ureteral epithelial cells. (e,f) Fluorescence microscopic image of (e) purified CK+DAPI+CD13+CD10+ cells, and (f) purified CK+DAPI+CD326+CD227+ cells (blue = DAPI, green = cytokeratin). (g–j) Electron microscopic images of purified cytokeratin+CD227+ cells using a cell sorter. (g and detail h) Apoptotic cell with characteristic marginalization of condensed chromatin (yellow arrows in h) and intact plasma membrane. (i and j) Necrotic cells with chromatin clumping (red arrow in j) and loss of plasma membrane integrity as distinguishing feature from apoptosis (yellow arrow in j). TECs, tubular epithelial cells.

Article Snippet: The complete staining protocol included pan-cytokeratin-APC (clone: REA831; isotype: recombinant human IgG1) or, exclusively for fluorescence microscopy, pan-cytokeratin-FITC (clone: CK3-6H5; isotype: mouse IgG1), CD13-PE (clone: REA263; isotype: recombinant human IgG1), CD10-PE-Vio770 (clone: 97C5; isotype: mouse IgG1), CD326-PE (clone: HEA-125; isotype: mouse IgG1), and CD227-PE-Vio770 (clone: REA448; isotype: recombinant human IgG1) (all Miltenyi Biotec GmbH, Bergisch Gladbach, Germany).

Techniques: Flow Cytometry, Staining, Fluorescence, Purification, Clinical Proteomics, Membrane

Journal: bioRxiv

Article Title: Castration-mediated IL-8 Promotes Myeloid Infiltration and Prostate Cancer Progression

doi: 10.1101/651083

Figure Lengend Snippet: a, Representative images of Cxcl15 fluorescent detection (murine homologue of IL-8) in Myc-Cap tumors. Tumors were harvested when volumes reached ∼500mm (CS group), 7 days after androgen-deprivation (ADT), or at the time of castration-resistance (CR) and hybridized with CF568-labeled probe sets (white) to Cxcl15 , CF640-labeled anti-PanCK antibody (red), and CF488-labeled anti-CD45 antibody (green). Nuclei counterstained with DAPI (blue). Repeated × 3. b, Gene and protein expression of Cxcl15 in MCRedAL cells of indicated tumor samples by qRT-PCR and ELISA, respectively (n = 3 per group, repeated × 2). c, qRT-PCR quantification of IL-8 in human AR positive castration-sensitive cells (CS: LNCaP, LAPC4, and VCaP) and their castration-resistant counterparts (CR: LNCaP-abl, LAPC4-CR, and VCaP-CR), replicate numbers as in b . d, IL-8 protein expression in the isogenic cell pairs from c quantified by ELISA, replicate numbers as in c . e, Representative images of Cxcl15 fluorescent detection in benign murine prostate tissue samples from castration-sensitive (CS), androgen-deprivation treated (ADT), and ADT-treated mice that received testosterone repletion (ADT + T). Tissue sections hybridized with CF568-labeled probe sets (white) to Cxcl15 , and CF640-labeled anti-PanCK antibody (red). Nuclei were counterstained with DAPI (blue). Repeated × 3. f, qRT-PCR analysis of Cxcl15 expression in prostate luminal epithelial cells from indicated treatment groups (n = 3 per group). Prostate luminal epithelial cells were isolated based on their GFP + CD49f int CD24 + CD45 - F4/80 - CD11b - expression by flow sorting into Trizol LS. g, Expression of IL-8 in human prostate epithelial cells micro-dissected from patients in a clinical trial ( NCT00161486 ) receiving placebo, androgen-deprivation treatment (ADT), or ADT plus testosterone repletion (ADT + T). Z-score values of microarray transcripts from benign prostate biopsies were normalized to placebo samples (n = 4 per group; GSE8466). h, Expression of IL-8 in human prostate cancer epithelial cells micro-dissected from untreated or ADT-treated ( NCT01696877 ; n = 8 per group) patients as determined by qRT-PCR. RISH images are at 60X magnification; scale bar = 100 μm. Gene expression levels were normalized to the mean ΔCT level in samples from CS, untreated or placebo groups. For b-g , unpaired t-tests were performed; for h a Mann-Whitney U test was used due to the non-normal data distribution observed. p -values ≤ 0.05 (*) and 0.01 (**); p -values ≥ 0.05 (ns) shown. The range in box and whiskers plots shows min and max values such that all data are included.

Article Snippet: Primary antibody for PanCK was diluted 1/400 in renaissance background reducing diluent (Biocare Medical) and incubated overnight at 4 °C.

Techniques: Labeling, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Isolation, Microarray, MANN-WHITNEY

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Spatial transcriptomics reveals prognostically LYZ + fibroblasts and colocalization with FN1 + macrophages in diffuse large B-cell lymphoma

doi: 10.1007/s00262-025-03968-7

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Mouse anti-human pan Cytokeratin IgG antibody , GB122053 , Servicebio.

Techniques: Formalin-fixed Paraffin-Embedded, Gene Expression, RNA Sequencing, Sequencing, Biomarker Discovery, Clinical Proteomics, Microarray, Control, Immunohistochemistry, Microscopy, Immunofluorescence, Software