Journal: Cell Death & Disease

Article Title: CXCR6+ T cells promote apoptosis and necroptosis in proximal tubules during AKI-to-CKD transition

doi: 10.1038/s41419-026-08644-x

Figure Lengend Snippet: A – D Wild-type and Cxcr6 -/- mice were subjected unilateral ischemia/reperfusion injury (IRI). The injured (IRI) kidneys were harvested 14 days after U-IRI. A Kidney sections were immunofluorescence-stained with KIM-1 (green), VCAM-1 (red), and DAPI (blue). Original magnification, ×40. Scale bar: 200 μm. B Fluorescence positivity was quantified using ImageJ. n = 4 CTRL kidneys/genotype and n = 8 IRI kidneys/genotype. Two-way ANOVA (injury and genotype interaction): P = 0.0623 (KIM-1) and P = 0.0060 (VCAM-1). * P < 0.05, *** P < 0.001, and **** P < 0.0001 by Tukey’s multiple comparison. ns not statistically significant. C Quantitative PCR for Havcr1 and Vcam1 was performed on whole-kidney mRNA. n = 8 mice/genotype. Two-way ANOVA (injury and genotype interaction): P < 0.0001 ( Havcr1 ) and P = 0.0006 ( Vcam1 ). **** P < 0.0001 by Tukey’s multiple comparison. ns not statistically significant. D Kidney sections were immune-stained with LTL (dark gray) (representative images shown), and LTL positive area was quantified in ( E ). F Kidney sections were stained for H&E (representative images shown), and cast area was quantified in ( G ). Green arrows in D indicate LTL-positive tubule. Red arrows in D and black arrows in F indicate tubular cast. Scale bar: 200 μm. n = 8 IRI kidneys/genotype. * P < 0.05 and ** P < 0.01 by unpaired t test. H Kidney sections were immunohistochemistry-stained for SOX9 (representative images shown). Scale bar: 50 μm. Arrows indicate SOX9-positive tubular cells. I Quantitation of SOX9-positive tubular cell per high power field (HPF), as in ( H ). n = 4 CTRL kidneys/genotype and n = 8 IRI kidneys/genotype. Two-way ANOVA (genotype and injury interaction): P = 0.0032. * P < 0.05 and **** P < 0.0001 by Tukey’s multiple comparison. ns not statistically significant. J Quantitative PCR for Sox9 was performed on whole-kidney mRNA. n = 8 mice/genotype. Two-way ANOVA (injury and genotype interaction): P = 0.0151. ** P < 0.01 and **** P < 0.0001 by Tukey’s multiple comparison. ns not statistically significant. K Scheme of experimental design. Wild-type and Cxcr6 −/− mice were subjected U-IRI, followed by contralateral nephrectomy (CL-NX) on day 14 after U-IRI. The blood was withdrawn for blood urea nitrogen (BUN in L ) and serum creatinine (SCr in M ) at the indicated time points. n = 14, 15 mice/genotype. Two-way ANOVA (injury and genotype interaction): P < 0.0001 (BUN and SCr) by two-way ANOVA. * P < 0.05, *** P < 0.001, and **** P < 0.0001 by Tukey’s multiple comparison. ns not statistically significant.

Article Snippet: Cleaved caspase 3, phospho-MLKL, KIM-1, VCAM-1, LTL, CXCR6, and F4/80 were detected by IF using primary antibodies against, cleaved caspase 3 (clone: 5A1E, Cell Signaling Technology), MLKL (phospho S345) (#37333, Cell Signaling Technology), KIM-1 (#AF1817, Novus Biologicals), VCAM-1 (#32653, Cell Signaling Technology), Ki67 (#12202, Cell Signaling Technology), TUNEL-fluorescein (#11684795910, Roche), LTL-fluorescein (# L32480 , Thermo Fisher Scientific), CXCR6 (#NLS1102, Novus Biologicals), and F4/80 (#MCA497, Bio-Rad), respectively.

Techniques: Immunofluorescence, Staining, Fluorescence, Comparison, Real-time Polymerase Chain Reaction, Immunohistochemistry, Quantitation Assay

Journal: Nature communications

Article Title: Single-cell and spatial transcriptome analyses reveal tertiary lymphoid structures linked to tumour progression and immunotherapy response in nasopharyngeal carcinoma.

doi: 10.1038/s41467-024-52153-4

Figure Lengend Snippet: Fig. 6 | Co-culture assays of CAFs and B cells. a Schematic diagram showing the B cell adhesion assay (left panel). Bar plots showing the capability of B cell adhesion to CAFs (right panel; n = 3). B cells were cocultured with either VCAM1- CAFs, VCAM1+ CAFs, or VCAM1+ CAFs in combination with a VCAM1 neutralising antibody. The relative number (OD value) of B cells bound to CAFs were quantified using CCK8 assays. b Bar plot showing the relative expression level of CXCL13 in fibro- blasts (left panel; n = 3). Fibroblasts were transduced with an empty vector or CXCL13 plasmid. Bar plot illustrates the migration capabilities of B cells under various co-culture conditions with fibroblasts (right panel; n = 11). The B cells were co-cultured with fibroblasts overexpressing either an empty vector (Con_EV), CXCL13 alone, or CXCL13 in combination with a CXCL13-specific neutralising antibody. c Schematic diagram showing the B cell co-culture assay (left panel). Bar

Article Snippet: Then, VCAM1+ or VCAM1- CAFs (2 × 104 per well) were seeded on a 96-well plate foroneday.On the secondday, CAFswere incubated with the presence or absence of neutralising antibody anti-VCAM1 Antibody (30μg/ml each, R&D systems, BBA5) for one hour prior to the adhesion assay.

Techniques: Co-Culture Assay, Cell Adhesion Assay, Expressing, Transduction, Plasmid Preparation, Migration, Cell Culture, Co-culture Assay