Journal: bioRxiv

Article Title: Characterization of bronchovascular-bundle mesenchymal stromal cells regulating antibody-secreting cell niche in rejecting lung allografts

doi: 10.1101/2025.02.16.638532

Figure Lengend Snippet: ( A ) FACS–sorted cell populations from adult mouse lung were analyzed by real-time PCR. Values are represented as means ± SEM. *p<0.05, ****p<0.0001. One-way ANOVA; post hoc test: Bonferroni’s. ( B ) Uniform manifold approximation and projection (UMAP) plot from scRNA-Seq on CD45 − CD31 − cell populations sorted from adult mouse lungs . ( C ) UMAP plots of Cxcl12 and ll6, with coordinates same as in B , but the color intensity indicates gene expression level within each cell. ( D ) Violin plots show expression intensity of Cxcl12 and Il6 in each of the 14 clusters. ( E ) Correlation plot showing Foxf1_MC cluster distinctly expressing Cxcl12 and Il6 gene expression. ( F ) Radar plot representing differentially expressed key genes marking the signature of each lung mesenchymal clusters. ( G ) Cxcl12 CreERT2 Rosa26 tdTomato adult mouse lung sections were immunolabeled for CXCL12 (red), mesenchymal αSMA (green) and epithelial EpCAM (white). Bronchovascular field shows presence of peri-bronchial and perivascular mesenchymal cells staining positive for CXCL12. n=3 per group. Original magnification: 200x; Scale bar = 50 µm. Abbreviations: AW, airway; V, vessel.

Article Snippet: CXCL12 levels were measured using the Human CXCL12/SDF-1⍺ Quantikine ELISA Kit (DSA00, R&D Systems) according to the manufacturer’s protocol.

Techniques: Real-time Polymerase Chain Reaction, Gene Expression, Expressing, Immunolabeling, Staining

Journal: bioRxiv

Article Title: Characterization of bronchovascular-bundle mesenchymal stromal cells regulating antibody-secreting cell niche in rejecting lung allografts

doi: 10.1101/2025.02.16.638532

Figure Lengend Snippet: ( A ) Flow cytometry to sort cell populations from Cxcl12 CreERT2-tdTomato mice that are CD45 − /CD31 − /EpCAM − /PDGFRα + MCs that are CXCL12(+) or CXCL12(−). ( B ) Cxcl12 and Il6 mRNA expression analyzed by real-time PCR of sorted populations from A. ( C ) mRNA expression of pro-fibrotic genes by real-time PCR of sorted populations from A. Values are represented as means ± SEM. *p<0.05, **p<0.01, ****p<0.0001 . One-way ANOVA; post hoc test: Bonferroni’s. ( C ) Immunofluorescent imaging of representative tissue section from Cxcl12 CreERT2-tdTomato mice in the pleural region showing CXCL12+ MCs (red). ( D ) Immunofluorescent imaging of representative tissue section from day 28 murine RAS allografts in the pleural region showing CD138 (red) overlapping with Ig deposition (green) and B220 (white). C-D. n=3. Original magnification = 200x. Scale bar = 50 μm.

Article Snippet: CXCL12 levels were measured using the Human CXCL12/SDF-1⍺ Quantikine ELISA Kit (DSA00, R&D Systems) according to the manufacturer’s protocol.

Techniques: Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction, Imaging

Journal: bioRxiv

Article Title: Characterization of bronchovascular-bundle mesenchymal stromal cells regulating antibody-secreting cell niche in rejecting lung allografts

doi: 10.1101/2025.02.16.638532

Figure Lengend Snippet: ( A ) Cxcl12 and Il6 mRNA expression analyzed by real-time PCR in FACS-sorted cell populations from an isograft and day 28 RAS allografts. Values are represented as means ± SEM. *p<0.05, **p<0.01, ****p<0.0001 . One-way ANOVA; post hoc test: Bonferroni’s. ( B ) Schema for the generation of Cxcl12 CreERT2-tdTomato donor mice and lung transplant experiments. TAM, tamoxifen. ( C ) Representative immunofluorescence imaging of Cxcl12-tdTomato Red + MCs in n=3 RAS allografts. Scale bars: 100 μm (zoom, cropped: 50 μm).

Article Snippet: CXCL12 levels were measured using the Human CXCL12/SDF-1⍺ Quantikine ELISA Kit (DSA00, R&D Systems) according to the manufacturer’s protocol.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Immunofluorescence, Imaging

Journal: bioRxiv

Article Title: Characterization of bronchovascular-bundle mesenchymal stromal cells regulating antibody-secreting cell niche in rejecting lung allografts

doi: 10.1101/2025.02.16.638532

Figure Lengend Snippet: ( A, B ) CXCL12 mRNA expression analyzed by real-time PCR in MCs derived from lung transplant patients diagnosed with (CLAD) or without CLAD (non-CLAD) ( A ) and in non-CLAD MCs treated with TGF-β (2 ng/mL) and LPA (10 μM) ( B ). ( C ) non-CLAD MCs were treated with vehicle, IL-6 (50 ng/mL), sIL-6R (200 ng/mL) or the combination of IL-6 and sIL-6R for 30 minutes. Expression of CXCL12 mRNA (real-time PCR) and protein (ELISA) in MCs and conditioned media, respectively. ( D-F ) non-CLAD MCs were pre-treated with JAK1/2 pan-inhibitor, Ruxolitinib (RUX), or JAK3 inhibitor, WHI-P154 (WHI), and the transcription inhibitor, Actinomycin D (ActD) and then stimulated with IL-6 and sIL-6R. D. CXCL12 and IL6 mRNA expression in MCs (real-time PCR) E. CXCL12 protein expression analyzed by ELISA in the conditioned media. F. Representative western blot of STAT3-Tyr705 phosphorylation. ( G, H ) non-CLAD MCs were transfected with lentiviral vectors that allowed constitutional activation of STAT3 (CA- STAT3 ) or scrambled controls. G. STAT3 and IL6 mRNA expression analyzed by real-time PCR. H. CXCL12 mRNA and protein expression in MCs (real-time PCR) and conditioned media (ELISA), respectively. I. non-CLAD MCs were transfected with scrambled control siRNA or sequences specific to CXCL12 , and then stimulated with IL-6 and IL-6R for 48h. Conditioned media was used to stimulate 0.5 million splenocytes in the upper chamber to migrate through the membrane. The number of CD3 − CD19 + B lymphocytes in the lower chamber was measured by FACS analyses. Values are represented as means ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 . One-way ANOVA; post hoc test: Bonferroni’s ( A-E , G-I ).

Article Snippet: CXCL12 levels were measured using the Human CXCL12/SDF-1⍺ Quantikine ELISA Kit (DSA00, R&D Systems) according to the manufacturer’s protocol.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Derivative Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Phospho-proteomics, Transfection, Activation Assay, Control, Membrane

Journal: bioRxiv

Article Title: Characterization of bronchovascular-bundle mesenchymal stromal cells regulating antibody-secreting cell niche in rejecting lung allografts

doi: 10.1101/2025.02.16.638532

Figure Lengend Snippet: ( A ) Experimental schematic. B6D2F1/J donor lungs were transplanted into C57BL/6J recipient mice, followed by treatment with Olamkicept (2.5 mg/kg/week; intraperitoneal injections) between days 3 and 28. ( B ) CXCL12 protein concentration by ELISA in lung homogenates from A . ( C , D ) Isografts, or RAS allografts treated with or without Olamkicept or Bortezomib were sacrificed at day 28. Serum was analyzed for IgG ( C ) and IgM (C) levels by ELISA. ( E ) FACS analyses of antibody secreting cells (ASCs). (F) Immunofluorescent staining of representative tissue sections showing antibody secreting cells marked by B220/CD138/IgG and airway epithelial integrity marked by CCSP. Representative sections stained for Movat Pentachrome. n=3. Original magnification = 200x. Scale bar = 50 μm. ( G ) Quantitative assessment of fibrosis by measuring hydroxyproline content in graft lung homogenates. Values are represented as means ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. One-way ANOVA; post hoc test: Bonferroni’s ( B-E , G ).

Article Snippet: CXCL12 levels were measured using the Human CXCL12/SDF-1⍺ Quantikine ELISA Kit (DSA00, R&D Systems) according to the manufacturer’s protocol.

Techniques: Protein Concentration, Enzyme-linked Immunosorbent Assay, Staining

Journal: The Prostate

Article Title: Pilot and feasibility study of serum chemokines as markers to distinguish prostatic disease in men with low total serum PSA.

doi: 10.1002/pros.20717

Figure Lengend Snippet: Fig. 1. A:LogserumCXCL5orCXCL12(ng/ml)differsdependingonprostatediseasestatus.ELISA-derivedvaluesforserumCXCL12formen withoutevidenceofprostaticdisease(whitetriangles),menwithoutcancerbutwithhistologicalprostatitis(gray triangles)andmenwithpros- tatecancer(blacktriangles),aswellasthelogserumCXCL5formenwithoutevidenceofprostaticdisease(whitediamonds),menwithoutcancer butwithhistologicalprostatitis (graydiamonds), andmenwithprostate cancer (blackdiamonds) are shown on a logarithmic scale. Significant differences (P < 0.050) are indicatedby *, trends (0.065 < P < 0.050) by #.B: Log serum CXCL5 (ng/ml) relevant to prostate volume.ELISA- derivedvalues for serumCXCL5 formenwithlow volume(37.5g)(circles)or high-volume(>37.5g)(diamonds) prostateswithoutcanceror histologicalprostatitis (white), withoutcancer butwithhistologicalprostatitis (gray), or with cancer (black) are shown on a logarithmic scale Differencesbetweengroupsthatachievedstatisticalsignificance(P < 0.05)areindicatedby*;trendsby#.Thedatarepresentedherearedrawn fromTable II.C: Log serum CXCL12 (ng/ml) relevant to prostate volume.ELISA-derived values for serum CXCL12 for men with low volume (37.5 g) (circles) or high-volume (>37.5 g) (diamonds) prostates without cancer or histological prostatitis (white), without cancer but with histological prostatitis (gray), or with cancer (black) are shown on a logarithmic scale. Differences between groups that achieved statistical significance (P < 0.05) is indicated by *; trends by #.The data represented here is drawn fromTable II.D: Log chemokine values in serum and plasma.ELISA-derivedvalues for CXCL12 (left) or CXCL5 (right) from serum (diamonds) orplasma (squares) are shown on alogarithmic scale for6of the51patientsexaminedinthisstudy.

Article Snippet: Circulating serum CXCL5 (ENA-78) or CXCL12 (SDF-1) chemokine levels were assessed using 50 ul frozen serum or plasma per direct sandwich ELISA in duplicate using the Human CXCL5/ENA-78 DuoSet kit DY254 or theHumanCXCL12/SDF-1 alpha capture antibody MAB350, detection antibody BAF310, and standard 350-NS ELISA reagents (R&D Systems).

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay, Clinical Proteomics

Journal: BMC Cancer

Article Title: CXCL12 expression by healthy and malignant ovarian epithelial cells

doi: 10.1186/1471-2407-11-97

Figure Lengend Snippet: CXCL12 expression in healthy and malignant ovaries . (A) Healthy ovary (i), CXCL12 immunoreactivity in OSE (inset is the outlined region on the tissue specimen containing surface epithelium, × 40), faint staining in the stroma and no signal in follicles and oocytes (arrow) (× 20); fallopian tube (ii), CXCL12 immunoreactivity in cells of the epithelium (× 40). (B) Serous (i) and mucinous (ii) benign epithelial ovarian tumors, CXCL12 immunoreactivity in proliferating epithelial cells (× 40). (C) Serous (i and iii) and mucinous (ii and iv) borderline epithelial ovarian tumors with low (CXCL12 low , i and ii) or high (CXCL12 high , iii and iv) levels of CXCL12 staining (× 40). (D) Malignant epithelial ovarian tumors: serous (i), mucinous (ii), clear-cell (iii) and endometrioid (iv), CXCL12 immunoreactivity in epithelial cells is confined to the cytoplasm, with frequent strong staining of the membrane (arrows), no staining in the nuclei of tumor cells or in the stroma (× 40). (E) Cytocentrifuged CD326 + epithelial (i) and CD326 - non epithelial (ii) cells isolated from malignant ascites collected from a patient diagnosed with invasive EOC, CXCL12 is detected only in CD326 + cells (× 40). (F) Non epithelial ovarian tumors: granulosa tumor (i) and dysgerminoma with characteristic morphological features, i.e. Exner bodies (arrow) (ii), absence of CXCL12 immunostaining from both tumors (× 40). No labeling was detected when the K15C anti-CXCL12 mAb was omitted or a 100-fold molar excess of recombinant CXCL12 was added to the mAb before incubation with tissues.

Article Snippet: For comparisons of CXCL12 mRNA levels in EOC samples, we used unpaired two-tailed Student's t tests (Prism software, GraphPad).

Techniques: Expressing, Staining, Membrane, Isolation, Immunostaining, Labeling, Recombinant, Incubation

Journal: BMC Cancer

Article Title: CXCL12 expression by healthy and malignant ovarian epithelial cells

doi: 10.1186/1471-2407-11-97

Figure Lengend Snippet: Steady-state levels of CXCL12 transcripts in healthy and malignant ovarian tissues . (A) CXCL12 mRNA was detected by conventional PCR and was of the expected size (417 bp) in healthy ovaries (faint signal), benign and invasive ovarian tumors (strong signal) and in CD326 + epithelial cell-enriched malignant ascites samples. By contrast, CXCL12 transcripts were absent from CD326 - non epithelial cells. The results presented are from one experiment representative of three carried out. The white vertical line separates lanes not run on the same gel. (B) CXCL12 mRNA levels were quantified by real-time PCR and are expressed as CXCL12 / β-actin ratios. The diagram shows the distribution of values and means for EOC samples identified as CXCL12 high and CXCL12 low/moderate . Each symbol represents an individual sample run in duplicate. The P value presented is that for a two-tailed Student's t test.

Article Snippet: For comparisons of CXCL12 mRNA levels in EOC samples, we used unpaired two-tailed Student's t tests (Prism software, GraphPad).

Techniques: Real-time Polymerase Chain Reaction, Two Tailed Test

Journal: BMC Cancer

Article Title: CXCL12 expression by healthy and malignant ovarian epithelial cells

doi: 10.1186/1471-2407-11-97

Figure Lengend Snippet: Correlation of CXCL12 expression with clinical parameters

Article Snippet: For comparisons of CXCL12 mRNA levels in EOC samples, we used unpaired two-tailed Student's t tests (Prism software, GraphPad).

Techniques: Expressing

Journal: BMC Cancer

Article Title: CXCL12 expression by healthy and malignant ovarian epithelial cells

doi: 10.1186/1471-2407-11-97

Figure Lengend Snippet: Hazard ratios for OS and PFS of 183 patients with EOC after univariate COX regression analysis of CXCL12 abundance and clinical and pathologic features

Article Snippet: For comparisons of CXCL12 mRNA levels in EOC samples, we used unpaired two-tailed Student's t tests (Prism software, GraphPad).

Techniques:

Journal: BMC Cancer

Article Title: CXCL12 expression by healthy and malignant ovarian epithelial cells

doi: 10.1186/1471-2407-11-97

Figure Lengend Snippet: Overall survival and progression-free survival as a function of CXCL12 expression . Plots of Kaplan-Meier estimates for overall survival (A) and progression-free survival (B) of EOC patients with tumor tissues identified as CXCL12 low/moderate (n = 97, solid blue line) or CXCL12 high (n = 86, dotted red line). P values are those obtained in log-rank tests.

Article Snippet: For comparisons of CXCL12 mRNA levels in EOC samples, we used unpaired two-tailed Student's t tests (Prism software, GraphPad).

Techniques: Expressing

Journal: Science Advances

Article Title: Stromal-derived MAOB promotes prostate cancer growth and progression

doi: 10.1126/sciadv.adi4935

Figure Lengend Snippet: ( A ) GSEA plots of “chemokine signaling pathway” and “chemotaxis” gene sets enriched in MAOB-OE PrSC cells compared with controls. ( B ) ELISA of CXCL12 secretion in the culture media of control and MAOB-KD PrSC cells ( n = 3). ( C ) Representative IHC images and corresponding Pearson’s correlation analysis of stroma-expressed MAOB and CXCL12 protein levels in a PC TMA ( n = 37). Scale bars, 100 μm. ( D ) Pearson’s correlation analysis of MAOB and CXCL12 mRNA levels in patient-derived cultured prostatic stromal cells (left, n = 20) and laser-capture microdissected breast tumor stroma (right, n = 53) from GSE34312 and GSE9014 datasets, respectively. Statistical analysis was performed using one-way ANOVA with Dunnett’s test in (B). Data represent means ± SEM. ** P < 0.01.

Article Snippet: TGFβ1 (ELH-TGFb1-1) and CXCL12 (DLH-SDF1a-1) protein levels in cell culture media were quantified by enzyme-linked immunosorbent assay (ELISA) (RayBiotech).

Techniques: Chemotaxis Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Cell Culture

Journal: Science Advances

Article Title: Stromal-derived MAOB promotes prostate cancer growth and progression

doi: 10.1126/sciadv.adi4935

Figure Lengend Snippet: ( A ) Western blot of Twist1 in control and MAOB-manipulated PrSC cells upon NAC (5 mM, 48 hours) or H 2 O 2 (40 μM, 24 hours) treatment. ( B ) ELISA of CXCL12 secretion in culture media of control and MAOB-OE PrSC cells treated with TWIST1 siRNA or NAC (5 mM, 48 hours) ( n = 3). ( C ) qPCR of CXCL12 in indicated PrSC cells upon NAC treatment (5 mM, 48 hours) or Twist1/ TWIST1 siRNA expression ( n = 3). ( D and E ) Determination of CXCL12 mRNA (D) and 0.7-kb promoter activity (E) in PrSC cells upon Twist1 expression and/or TGFβ1 treatment (10 ng/ml, 12 hours) ( n = 3). ( F ) Schematic diagrams of WT and mutated CXCL12 E-box/SBE-Luc constructs and determination of their activities in PrSC cells upon Twist1 expression and/or TGFβ1 treatment (10 ng/ml, 12 hours) ( n = 3). ( G ) Representative proximity ligation assay staining and quantitation of indicated Twist1-Smad interactions by per-nucleus fluorescence intensity in control and MAOB-OE PrSC cells. Smad antibody incubation alone served as negative control. Numbers of nuclei included for comparisons between groups are denoted. Scale bars, 50 μm. ( H ) Co-IP assays of indicated Twist1-Smad interactions in PrSC cells with coexpression of Twist1 and individual Smads. Immunoglobulin G (IgG) was used in IP as negative control. Ten percent input was blotted as positive control. ( I ) ChIP analysis of chromatin from control and MAOB-OE PrSC cells precipitated with anti-Twist1, anti-Smad4, or a control IgG, followed by qPCR probing the E-box/SBE-centric CXCL12 promoter region ( n = 3). ( J ) ChIP analysis of chromatin from PrSC cells precipitated with anti-Smad4 antibody and then reprecipitated with anti-Twist1 or a control IgG (re-ChIP), followed by qPCR probing the E-box/SBE-encompassing CXCL12 promoter sequence ( n = 3). Statistical analysis was performed using one-way ANOVA with Tukey’s test. Data represent means ± SEM. * P < 0.05, ** P < 0.01; ns, not significant.

Article Snippet: TGFβ1 (ELH-TGFb1-1) and CXCL12 (DLH-SDF1a-1) protein levels in cell culture media were quantified by enzyme-linked immunosorbent assay (ELISA) (RayBiotech).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Activity Assay, Construct, Proximity Ligation Assay, Staining, Quantitation Assay, Fluorescence, Incubation, Negative Control, Co-Immunoprecipitation Assay, Positive Control, Sequencing

Journal: Science Advances

Article Title: Stromal-derived MAOB promotes prostate cancer growth and progression

doi: 10.1126/sciadv.adi4935

Figure Lengend Snippet: ( A ) Quantitation of C4-2 and PC-3 PC cells in monoculture and coculture with indicated PrSC fibroblasts upon anti-CXCL12 (0.1 μg/ml) antibody treatment ( n = 3). ( B ) Quantitation of PC cells in coculture with indicated PrSC cells treated with rCXCL12 protein (50 ng/ml; n = 3). ( C ) Western blot of CXCR4 and CXCR7 in control and CXCR4-KD/CXCR7-KD PC cells. ( D and E ) Quantitation of control and CXCR4-KD/CXCR7-KD PC cells in coculture with indicated PrSC cells treated without (D) or with (E) rCXCL12 (50 ng/ml; n = 3). ( F ) Quantitation of PC cells in coculture with indicated PrSC cells treated with 10 nM AMD3100 ( n = 3). ( G ) Representative images and quantitation of PC-3 cell migration and invasion in coculture with indicated PrSC cells upon treatment with anti-CXCL12 antibody (0.1 μg/ml) or 10 nM AMD3100 ( n = 3). Scale bars, 200 μm. ( H ) Representative images and quantitation of PC cell migration in coculture with indicated PrSC cells treated with rCXCL12 (50 ng/ml; n = 3). Scale bars, 100 μm. ( I ) Phospho-antibody array analysis of PC-3 cells treated with indicated PrSC cell CM. All phosphoprotein levels were normalized to their total forms from a single array with six replicate spots, with significantly activated phosphoproteins (fold change > 1.5, P < 0.05) denoted. ( J ) Western blot of p-Src and p-JNK in control and CXCR4-KD PC-3 cells treated with indicated PrSC cell CM. ( K ) Western blot of p-Src and p-JNK in PC-3 cells treated with indicated PrSC cell CM plus rCXCL12 (50 ng/ml). ( L ) Quantitation of PC cells in coculture with indicated PrSC cells following pretreatment with 40 nM Src inhibitor 1 or 10 μM SP600125 for 24 hours ( n = 3). Statistical analysis was performed using one-way ANOVA with Tukey’s test. Data represent means ± SEM. * P < 0.05, ** P < 0.01; ns, not significant.

Article Snippet: TGFβ1 (ELH-TGFb1-1) and CXCL12 (DLH-SDF1a-1) protein levels in cell culture media were quantified by enzyme-linked immunosorbent assay (ELISA) (RayBiotech).

Techniques: Quantitation Assay, Western Blot, Migration, Ab Array

Journal: Science Advances

Article Title: Stromal-derived MAOB promotes prostate cancer growth and progression

doi: 10.1126/sciadv.adi4935

Figure Lengend Snippet: ( A ) Quantitation of C4-2 and PC-3 cell proliferation in monoculture and coculture with control and MAOB-OE PrSC cells upon selegiline treatment (10 nM, 72 hours) ( n = 3). ( B ) BLI-based growth curves of Luc-tagged PC-3 tumors grown in the prostates of male NSG mice treated with selegiline at various doses (0.5, 2, and 10 mg/kg) or saline as a vehicle ( n = 5 per group). ( C ) BLI images of mice from each group at the end point. ( D ) Determination of tumor weights ( n = 5). ( E ) Determination of MAOA and MAOB enzymatic activities in mouse liver tissue from each group at the end point ( n = 3). ( F ) Representative images of H&E and IHC staining of tumor-expressed Ki-67, p-Src, and p-JNK and stroma-expressed αSMA and CXCL12 and their quantitation in tumor samples from each group ( n = 5). Scale bars, 100 μm. ( G ) Mouse body weights determined weekly ( n = 5). ( H ) Representative H&E images of mouse liver and kidney tissue from each group. Scale bars, 100 μm. ( I to L ) ELISA of ALT (I), AST (J), BUN (K), and creatinine (L) in mouse sera at the end point ( n = 5). ( M ) Schematic depicting stromal-derived MAOB activation of paracrine CXCL12-CXCR4/Src/JNK signaling through interplay between ROS-dependent Twist1 (via a HIF1α/VEGF-A/AKT/FOXO1 pathway) and TGFβ1/Smads to promote stromal-epithelial interactions for PC growth and progression. Statistical analysis was performed using one-way ANOVA with Tukey’s test in (A), (D) to (F), and (I) to (L) and two-way ANOVA with Tukey’s test in (B) and (G). Data represent means ± SEM. * P < 0.05, ** P < 0.01; ns, not significant.

Article Snippet: TGFβ1 (ELH-TGFb1-1) and CXCL12 (DLH-SDF1a-1) protein levels in cell culture media were quantified by enzyme-linked immunosorbent assay (ELISA) (RayBiotech).

Techniques: Quantitation Assay, Saline, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Derivative Assay, Activation Assay