Journal: Aging Cell

Article Title: Single‐Cell Sequencing Reveals That CD4 + T Cells Eliminate Senescent Prostate Epithelium to Delay Progression of Benign Prostatic Hyperplasia

doi: 10.1111/acel.70180

Figure Lengend Snippet: Laser capture microdissection was used to isolate senescent and non‐senescent epithelial cells in BPH for transcriptomic and pathway analysis. (a) Schematic workflow showing the isolation of paired senescent and non‐senescent epithelial cells from BPH tissue using serial sectioning, SA‐β‐gal staining, and laser capture microdissection (LCM) for downstream transcriptomic analysis. (b) Volcano plot of differentially expressed genes (DEGs) between senescent and non‐senescent epithelial cells. CXCL13 is significantly upregulated in senescent cells. (c) Gene set enrichment analysis (GSEA) reveals significant enrichment of the REACTOME_CELLULAR_SENESCENCE and REACTOME_SASP pathways in senescent epithelial cells, along with immune‐related pathways including GSE26928_NAIVE_VS_CXCR5_POS_CD4_TCELL_DN and GOBP_NEGATIVE_REGULATION_OF_T_CELL_MEDIATED_IMMUNITY, suggesting immune activation and chemotactic signaling. (d) Heatmap displaying upregulation of key senescence‐ and inflammation‐associated genes such as TP53, CDKN1C (p57), SERPINE1, and other SASP factors in senescent epithelial cells. (e) MSigDB GO enrichment analysis showing overrepresentation of pathways related to senescence, lymphocyte activation, CD4 + T cell proliferation, and leukocyte chemotaxis. (f) Volcano plot showing transcriptomic differences between prostate tissues from individuals aged 70–79 and 20–29. CXCL13 is notably upregulated in the elderly group. (g) GSEA plots showing enrichment of FRIDMAN_SENESCENCE_UP, GOBP_LEUKOCYTE_CHEMOTAXIS, GOBP_POSITIVE_REGULATION_OF_T_CELL_PROLIFERATION, and GSE28726_NAIVE_VS_ACTIVATED_CD4_TCELL_DN in the older cohort, indicating enhanced senescence and T cell activation in aged prostate tissue. (h) MSigDB enrichment analysis further confirms significant immune activation in aged prostates, highlighting pathways involving CD4 + T cell recruitment and chemokine activity. (i) Venn diagram identifying CXCL13 as the only gene consistently upregulated in both senescent epithelial cells and aged prostates (70–79 vs. 20–29), emphasizing its conserved role in senescence and immune crosstalk. (j) Independent analysis using the ADEIP dataset confirms age‐associated upregulation of CXCL13, supporting its pivotal involvement in prostate aging and BPH development.

Article Snippet: Recombinant murine CXCL13 (1 μg/day; HY‐ P77911 , MedChemExpress) was administered intraperitoneally on days 1, 5, 8, 12, 15, and 19.

Techniques: Laser Capture Microdissection, Isolation, Staining, Activation Assay, Chemotaxis Assay, Activity Assay

Journal: Aging Cell

Article Title: Single‐Cell Sequencing Reveals That CD4 + T Cells Eliminate Senescent Prostate Epithelium to Delay Progression of Benign Prostatic Hyperplasia

doi: 10.1111/acel.70180

Figure Lengend Snippet: Immunohistochemical analysis of CXCL13, p16, and CD4 in BPH tissues reveals an age‐dependent association between senescent cells and immune infiltration. (a) Representative immunohistochemical (IHC) images of BPH tissue sections stained for CXCL13, p16, and CD4 show areas of high CXCL13 expression and CD4+ T cell infiltration in regions with elevated p16, a marker of cellular senescence. Images are presented at two magnifications: low magnification (scale bar = 200 μm, left) and high magnification (scale bar = 50 μm, right) to highlight areas of interest within the same field across markers. (b) Correlation analysis of IHC staining intensities (measured as optical density, OD, in units of 1 × 10 5 ) between CXCL13 and p16 (top), CD4 and CXCL13 (middle), and CD4 and p16 (bottom). Significant positive correlations were observed between CXCL13 and p16 ( R 2 = 0.2188, p = 0.012), CD4 and CXCL13 ( R 2 = 0.3617, p = 0.0007), and CD4 and p16 ( R 2 = 0.2561, p = 0.006), suggesting that senescent epithelial cells may recruit CD4+ T cells via CXCL13 signaling. (c–e) Scatter plots showing age‐related increases in staining intensities for CXCL13 (C; R 2 = 0.2905, p = 0.0031), p16 (D; R 2 = 0.2523, p = 0.0065), and CD4 (E; R 2 = 0.2253, p = 0.0107). These results indicate an age‐dependent rise in cellular senescence and CXCL13‐mediated recruitment of CD4+ T cells to senescent sites in BPH tissue. (f–i) Violin and bar plots displaying IHC scores for CXCL13 and CD4 stratified by p16 expression levels. (f) Violin plot showing significantly higher CXCL13 scores in high‐senescence (p16‐high) versus low‐senescence (p16‐low) groups ( p < 0.05). (g) Bar plot showing the distribution of CXCL13 staining intensities across p16 scores (1, 2, and 3), with higher p16 levels corresponding to stronger CXCL13 staining ( p < 0.001). (h) Violin plot showing significantly higher CD4 scores in high‐senescence groups ( p < 0.001). (i) Bar plot showing the distribution of CD4 staining intensities stratified by p16 scores, with increased CD4+ T cell infiltration correlating with higher p16 expression ( p < 0.001). Statistical significance: * p < 0.05, *** p < 0.001.

Article Snippet: Recombinant murine CXCL13 (1 μg/day; HY‐ P77911 , MedChemExpress) was administered intraperitoneally on days 1, 5, 8, 12, 15, and 19.

Techniques: Immunohistochemical staining, Staining, Expressing, Marker, Immunohistochemistry

Journal: Aging Cell

Article Title: Single‐Cell Sequencing Reveals That CD4 + T Cells Eliminate Senescent Prostate Epithelium to Delay Progression of Benign Prostatic Hyperplasia

doi: 10.1111/acel.70180

Figure Lengend Snippet: CXCL13‐CXCR5‐driven chemotaxis and HLA‐DR‐dependent apoptosis in the immune clearance of senescent BPH epithelial cells by CD4+ T cells. (a) Schematic of the transwell chemotaxis assay setup. CD4+ T cells were placed in the upper chamber, with conditioned media (CM) from senescent or normal BPH‐1 and PrEC epithelial cells in the lower chamber. CD4+ T cell migration toward senescent or normal epithelial CM was quantified after 12 h. (b, c) Fold change in CD4+ T cell migration toward conditioned media from senescent versus normal BPH‐1 (b) and PrEC (c) epithelial cells. Senescent CM significantly enhanced CD4+ T cell migration compared to normal CM. (d, e) CXCL13 expression levels in senescent and normal BPH‐1 (d) and PrEC (e) epithelial cells, as quantified by ELISA and qRT‐PCR. CXCL13 expression was significantly higher in senescent cells compared to normal cells. (f) Fold change in CD4+ T cell migration toward senescent BPH‐1 and PrEC CM, with and without anti‐CXCR5 or anti‐CXCL13 antibodies (1 μg/mL). The addition of anti‐CXCR5 or anti‐CXCL13 significantly reduced CD4+ T cell migration, demonstrating the involvement of the CXCL13‐CXCR5 axis in chemotaxis. (g) Volcano plot depicting differentially expressed genes between senescent and normal PrEC cells co‐cultured with CD4+ T cells at a 1:50 ratio. This plot highlights the upregulation of MHC‐II and SASP‐related genes, illustrating the significant changes in senescent PrEC's gene expression induced by T cell interactions. (h) Heatmap of key senescence and SASP markers in senescent versus normal epithelial cells, with significant upregulation of HLA‐DRB1, IL1A, IL1B, CXCL14, TNF, MMP13, and MMP14 in senescent cells, indicating the senescent phenotype. (i) GO enrichment analysis showing pathways related to chemokine signaling, MHC‐II antigen presentation, and T cell activation. Pathways like positive regulation of CD4+ T cell activation and T cell receptor signaling were enriched in senescent cells, supporting their role in immune interactions. (j) GSEA plots for selected pathways, including FRIDMAN_SENESCENCE_UP, REACTOME_MHC_CLASS_II_ANTIGEN_PRESENTATION, GOBP_REGULATION_OF_CD4_POSITIVE_ALPHA_BETA_T_CELL_ACTIVATION, and GOBP_POSITIVE_REGULATION_OF_EXECUTION_PHASE_OF_APOPTOSIS. These pathways are crucial in senescence, antigen presentation, and CD4+ T cell activation. (k) Flow cytometry plots showing apoptosis rates in BPH‐1 and PrEC cells (both normal and senescent) co‐cultured with CD4+ T cells at different ratios (0:1, 10:1, 50:1 + Iso, 50:1 + anti HLA‐DR). Apoptosis was measured by Annexin V‐FITC and PI staining. (l, m) Quantification of apoptosis percentages in BPH‐1 (L) and PrEC (M) epithelial cells. Apoptosis rates in senescent cells increased significantly with higher CD4+ T cell ratios. HLA‐DR inhibition at a 50:1 ratio significantly reduced apoptosis in senescent cells compared to isotype controls, highlighting the role of HLA‐DR‐mediated interactions in immune clearance. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant murine CXCL13 (1 μg/day; HY‐ P77911 , MedChemExpress) was administered intraperitoneally on days 1, 5, 8, 12, 15, and 19.

Techniques: Chemotaxis Assay, Migration, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Cell Culture, Gene Expression, Immunopeptidomics, Activation Assay, Flow Cytometry, Staining, Inhibition

Journal: Aging Cell

Article Title: Single‐Cell Sequencing Reveals That CD4 + T Cells Eliminate Senescent Prostate Epithelium to Delay Progression of Benign Prostatic Hyperplasia

doi: 10.1111/acel.70180

Figure Lengend Snippet: Effects of CXCL13 and CD4+ T cell depletion on prostate enlargement and senescence in a testosterone‐induced BPH model. (a) Experimental design for BPH induction in male BALB/c mice. Mice were divided into four groups: control ( n = 8), BPH ( n = 8), BPH + rmCXCL13 ( n = 8), and BPH + rmCXCL13 + anti‐CD4 ( n = 8). (b) Comparison of Prostate Index (PI) Across Groups: The PI was significantly higher in the BPH group than in the control group, confirming successful BPH induction. In the BPH + rmCXCL13 group, there was a notable reduction in PI, indicating that CXCL13 mitigates prostate enlargement. Conversely, the BPH + rmCXCL13 + anti‐CD4 group displayed a significant increase in PI relative to the BPH + rmCXCL13 group, suggesting that depletion of CD4+ T cells exacerbates CXCL13‐mitigated prostate hyperplasia. (c) Representative flow cytometry gating strategy and CD4+ T cell percentage within the CD3+ T cell population in prostate tissue across groups. (d) Quantification of CD4+/CD3+ T cell ratios across treatment groups. The BPH + rmCXCL13 group showed a significant increase in CD4+ T cell infiltration compared to the BPH group, indicating CXCL13‐mediated CD4+ T cell recruitment. In the BPH + rmCXCL13 + anti‐CD4 group, CD4+ T cells were significantly reduced, confirming effective CD4+ T cell depletion. (e) Immunohistochemical staining of prostate tissue for CXCL13, CD4, and p16 across treatment groups. CXCL13 expression increased from the control to the BPH and BPH + rmCXCL13 groups, with no significant difference between the BPH + rmCXCL13 and BPH + rmCXCL13 + anti‐CD4 groups. CD4 staining intensity was significantly higher in the BPH and BPH + rmCXCL13 groups but markedly reduced in the BPH + rmCXCL13 + anti‐CD4 group, confirming CD4+ T cell depletion. P16 staining, a marker of cellular senescence, was significantly elevated in the BPH group compared to the control. In the BPH + rmCXCL13 group, p16 expression was reduced but increased again in the BPH + rmCXCL13 + anti‐CD4 group. (f–h) Quantification of IHC scores for CXCL13, CD4, and p16 across groups, showing statistical comparisons. (i, k, m) Volcano plots of differential gene expression analyses for BPH vs. control (i), BPH + rmCXCL13 vs. BPH (k), and BPH + rmCXCL13 + anti‐CD4 vs. BPH + rmCXCL13 (m), highlighting significant changes in senescence‐related genes. (j, l, n) Gene Set Enrichment Analysis (GSEA) results for senescence and CD4+ T cell‐related gene sets. In BPH vs. control, FRIDMAN_SENESCENCE_UP, GSE6259_CD4_TCELL_VS_CD8_TCELL_UP, and GOBP_T_CELL_ACTIVATION_VIA_T_CELL_RECEPTOR_CONTACT_WITH_ANTIGEN_BOUND_TO_MHC_MOLECULE_ON_ANTIGEN_PRESENTING_CELL gene sets were significantly enriched in the BPH group. In BPH + rmCXCL13 vs. BPH, GOBP_CELLULAR_SENESCENCE, GSE2770_UNTREATED_VS_ACT_CD4_TCELL_48H_UP, and GSE13738_RESTING_VS_TCR_ACTIVATED_CD4_TCELL_UP gene sets were enriched in the BPH + rmCXCL13 group, with negative NES values indicating pathway reduction. In BPH + rmCXCL13 + anti‐CD4 vs. BPH + rmCXCL13, GOBP_CELLULAR_SENESCENCE, GSE6259_CD4_TCELL_VS_CD8_TCELL_DN, and GSE13738_RESTING_VS_TCR_ACTIVATED_CD4_TCELL_UP gene sets were enriched in the BPH + rmCXCL13 + anti‐CD4 group, indicating a potential regulatory role of CD4+ T cells in senescence signaling. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Recombinant murine CXCL13 (1 μg/day; HY‐ P77911 , MedChemExpress) was administered intraperitoneally on days 1, 5, 8, 12, 15, and 19.

Techniques: Control, Comparison, Flow Cytometry, Immunohistochemical staining, Staining, Expressing, Marker, Gene Expression, Activation Assay

Journal: Nature materials

Article Title: Colony stimulating factor-1 receptor is a central component of the foreign body response to biomaterial implants in rodents and non-human primates.

doi: 10.1038/nmat4866

Figure Lengend Snippet: Figure 5 | CSF1R-dependent macrophages recruit fibrosis-potentiating B cells via CXCL13. a, CSF1R inhibition prevents the entire (innate and adaptive) immune response to implanted biomaterials. Phase contrast images showing that fibrosis, as compared with wild-type (WT) vehicle-treated control, was partially eliminated by CXCL13 neutralization, and completely eliminated with continuous CSF1R inhibition (inh.; 160 mg kg−1 body weight GW2580 s.c., daily) over a 14-day implant period. Scale bars, 2,000 µm. Fibrosis was reduced with CXCL13 neutralization by the same extent as that of the B cell knockout (B KO) shown in Fig. 3a. b, Flow analysis for responding macrophages, neutrophils and B cells dissociated directly from spheres (100 µl material in all cases) 14 days post-intraperitoneal (i.p.) implant, showing partial loss of cell presence with CXCL13 neutralization, and complete loss following CSF1R inhibition. c, Bright-field images showing that fibrosis, compared with vehicle control, was completely eliminated by both macrophage depletion (−Macrophages) and CSF1R inhibition. Scale bar, 1,000 µm. d, Flow analysis for responding host innate immune macrophages and neutrophils dissociated directly from spheres (100 µl of each material) 14 days post-i.p. implantation, showing the loss of immune adhesion with loss of fibrosis due to either macrophage depletion (−Macrphages) or CSF1R inhibition. e, NanoString expression analysis for all known cytokine and cytokine receptors (see Supplementary Fig. 18 for complete data set, excerpted here), showing similar unique factors increased across all material (hydrogel alginate, ceramic glass, and polymer polystyrene (PS)) groups. f, Confocal for DAPI (cellular nuclei), macrophage marker CD68 (green), B cell marker CD19 (magenta), fibrosis-associated myofibroblast marker alpha smooth muscle actin (αSMactin, myofibroblasts, red), overlay, and bright-field imaging, showing that CXCL13 neutralization resulted in loss of B cell recruitment. Scale bar, 200 µm. g, qPCR expression analysis of α-SMactin directly on retrieved spheres from vehicle-treated (WT), B KO, and CXCL13-neutralized WT mice, plotted relative to expression levels on spheres from WT mice. Statistical analysis: one-way ANOVA with Bonferroni multiple comparison correction ∗∗∗p<0.0001, versus vehicle. Error bars, mean ± s.e.m. n=5 mice per group. Experiments were repeated at least 2–3 times.

Article Snippet: To neutralize secreted CXCL13, anti-CXCL13 antibody (mAB470, R&D Systems) was injected i.p. in sterile 1× PBS at a dose of 100 μg per mouse, once every 3 days, starting 3 days prior to implantation as well.

Techniques: Inhibition, Control, Neutralization, Knock-Out, Expressing, Polymer, Marker, Imaging, Comparison

Journal: Nature materials

Article Title: Colony stimulating factor-1 receptor is a central component of the foreign body response to biomaterial implants in rodents and non-human primates.

doi: 10.1038/nmat4866

Figure Lengend Snippet: Figure 6 | Essential fibrotic cascade players are also increased in non-human primates. SLG20 (0.5 mm) hydrogel spheres were implanted into either the intraperitoneal (i.p.) or subcutaneous (s.c.) dorsal region of cynomolgus monkeys and retrieved by laparoscopy-guided tissue excision or biopsy punch after 28 days (ref. 27). a, Laparoscopy images taken on days 0 and 28 (initial implantation and retrieval from the i.p. space). b, H&E- and Masson’s trichrome-stained histological sections of excised i.p. omentum tissue at 28 days for non-fibrosed fat-laden (No material, Mock) or heavily collagen-deposited and sphere-embedded (Implanted) omental tissue. Scale bars, 400 µm. c, Confocal staining showing DAPI (cellular nuclei), innate immune macrophage marker CD68 (green), and fibrosis-associated activated myofibroblast alpha smooth muscle actin (αSMactin) staining (red), showing cellular infiltration around and fibrosis deposition on an embedded 500 µm alginate sphere. Inset: additional confocal for CSF1R (green), showing positive staining on not only more distant macrophages but also material-proximal and fused foreign body giant cells; both ×20 magnification. White scale bars: both 200 µm for each respective image. d, Flow analysis showing similar host innate immune macrophage (CD68+CD11b+, top right quadrants) and remaining neutrophil/myeloid (CD68−CD11b+, bottom right quadrants) cells across C57BL/6 mice and cynomolgus monkeys, dissociated directly from fibrosed spheres and adjacent fibrosed omentum tissue (as percent composition) 28 days post-i.p. implantation. While the prominence of CD11b seems to be inverted in C57BL/6 mice versus cynomolgus monkeys, population response percentages are similar 28 days post-i.p. implantation. e, NanoString analysis for immune markers and cytokines, originally identified in C57BL/6 mice (note: CD66b is used here as a neutrophil marker, as Ly6g/Gr1 does not exist in NHPs or humans). Significant increases are observed for macrophage markers, as well as CSF1R and CXCL13 in both i.p. and s.c. implant sites, as compared with mock (saline-injected) controls (there was no difference between s.c. and i.p. mock controls). N=2 for i.p.-implanted groups; N=4 for s.c. treatment groups. These experiments were performed once for s.c. and twice for i.p. delivery.

Article Snippet: To neutralize secreted CXCL13, anti-CXCL13 antibody (mAB470, R&D Systems) was injected i.p. in sterile 1× PBS at a dose of 100 μg per mouse, once every 3 days, starting 3 days prior to implantation as well.

Techniques: Staining, Marker, Saline, Injection

Journal: Cancer immunology research

Article Title: Radiofrequency ablation remodels the tumor microenvironment and promotes neutrophil-mediated abscopal immunomodulation in pancreatic cancer

doi: 10.1158/2326-6066.CIR-22-0379

Figure Lengend Snippet: Tumor size was recorded 4 days before (Initial), right before (Pre) and 4 days after (Post) Sham or RFA treatment. Proteome arrays were performed in locally ablated tumors and serum of ablated mice and compared to Sham-treated mice (Control). A, Experimental design of RFA-treated mice. B, Growth curves show Control tumors (n = 8) significantly increased in size 4 days after treatment when compared to RFA-treated (n = 8) and non-RFA treated (n = 7) tumors. C, At the time of euthanization only Sham-treated tumors (n = 8) had significantly increased in size compared with pretreatment size; no difference in size was observed in RFA-treated (n = 8) and non-RFA treated (n = 7) tumors Pre and Post RFA. D, ImageJ quantification of necrosis, which was detected by H&E staining. RFA significantly increased necrosis on the RFA- and non-RFA-treated tumors compared to control Sham-treated tumors. E, Representative composite H&E staining of control, RFA, and non-RFA treated tumors showing necrotic areas inside dashed lines. F, ImageJ quantification showing RFA increased cleaved caspase 3+ cells in the RFA-treated and non-RFA treated tumors compared to control Sham-treated control tumors, as assessed by IHC. G, Representative IHC staining for cleaved caspase 3 in control, RFA, and non-RFA treated tumors. H, ImageJ quantification revealed RFA significantly increased the number of granzyme B+ cells in the RFA-treated tumors compared to controls and non-RFA treated tumors, as assessed by IHC. I, IHC staining for granzyme B in control, RFA, and non-RFA treated tumors. J, RFA-treated tumors (n = 3) presented increased expression of C5/C5a, IL-23 and CXCL12 compared to control (n = 2) tumor content. K, CXCL10, CXCL12, CXCL13 and TIMP-1 were significantly elevated in serum from RFA-treated (n = 4) mice compared to Sham-treated (n = 3) control serum. Time x Treatment comparisons were performed using Two-way ANOVA, treatment only comparisons by One-way ANOVA and proteome arrays were analyzed by multiple t test. Bar plots showing mean with SEM were used to represent data. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001; n.s., not significant. Scale bars are 50μM.

Article Snippet: CXCL13 ELISA Mouse CXCL13/BLC/BCA-1 Quantikine ELISA Kit assay was used to determine CXCL13 levels in mouse serum, splenocytes and tumors homogenates, following the manufacturer instructions (R&D Systems, cat #MCX130).

Techniques: Control, Staining, Immunohistochemistry, Expressing

Journal: Cancer immunology research

Article Title: Radiofrequency ablation remodels the tumor microenvironment and promotes neutrophil-mediated abscopal immunomodulation in pancreatic cancer

doi: 10.1158/2326-6066.CIR-22-0379

Figure Lengend Snippet: A-B, IMC analysis of tumors 4 days after Sham or RFA treatment revealed Ly6G+CD11b+CD44+ neutrophils are enriched in non-RFA treated tumors. C, Neighborhood analysis identified immune cells and markers with strong neutrophil co-localization. D, Cluster and Cell Phenotype information of Neighborhood analysis of IMC data. E, Experimental design for neutrophil depletion in vivo followed by RFA. F, RFA locally ablated tumors treated with IgG2a isotype control (VEH, n = 6) or anti-Ly6G (ND, n = 8) did not show differences in tumor size right before (Pre) and 4 days after (Post) RFA ablation. In non-RFA treated tumors, anti-Ly6G (ND, n = 8) treatment revealed an increase in tumor size Post RFA treatment when compared to IgG2a isotype control (VEH, n = 6) treated tumors. G, Neutrophil depletion (ND; anti-Ly6G treated group) did not alter αSMA staining, detected by IHC, in RFA-treated tumors when compared to RFA-treated tumors with IgG2a (VEH); on the contrary, neutrophil depletion (ND) revealed non-RFA treated tumors presented a significant increase in αSMA compared to control non-neutrophil depleted (VEH) group. H, Neutrophil depletion did not alter CD31 staining, detected by IHC, in any of the groups. I, Neutrophil-depleted RFA treated tumors presented a significant reduction in CXCL13 content compared to both VEH + RFA and non-RFA treated tumors when assayed using a cytokine array. No differences were found in non-RFA treated tumors between treatments. J, Neutrophil depletion presented a trend in reducing systemic CXCL13 levels in RFA treated mice. Tumor volume was analyzed by paired Student’s t test. Tumor chemokine levels were studied by Two-way ANOVA. IHC and serum protein expression levels were analyzed by unpaired Student’s t test. Bar plots indicate mean with SEM. *, P ≤ 0.05; **, P ≤ 0.01; ****, P ≤ 0.0001; n.s., not significant.

Article Snippet: CXCL13 ELISA Mouse CXCL13/BLC/BCA-1 Quantikine ELISA Kit assay was used to determine CXCL13 levels in mouse serum, splenocytes and tumors homogenates, following the manufacturer instructions (R&D Systems, cat #MCX130).

Techniques: In Vivo, Control, Staining, Expressing