Journal: PLoS ONE

Article Title: MMP-9 and CXCL8/IL-8 Are Potential Therapeutic Targets in Epidermolysis Bullosa Simplex

doi: 10.1371/journal.pone.0070123

Figure Lengend Snippet: A. SQRT-PCR shows increased matrix metalloproteinase mRNA expression in EBS-DM cell lines (n = 4). B. MMP-9 ELISA of 48-h-conditioned cell culture supernatant shows 2-fold upregulation of MMP-9 in KEB-7 and 46-fold upregulation in EBDM-1 at the protein level (n = 4). C. MMP-9 levels were highly increased in EBS patients blister fluids compared to healthy controls (n = 3 to n = 4). The numbers correlate with . Student`s t -test was performed with p values: * ≤0.05, ** ≤0.01, *** ≤0.005, ΔΔ≤0.0005, ΔΔΔ≤0.0001. (In 2C, the Student’s t -test compared the entire patient group to the entire control group).

Article Snippet: Protein levels of KLK5, MMP7, MMP9, CXCL1, CXCL8/IL-8, CXCL11 and CXCL14 were determined in 48-h-conditioned cell culture supernatant and in patients blister fluids by using Quantikine® ELISA (Human KLK5, # DKK500, R&D Systems; Human MMP7, # DMP700, R&D Systems; Human MMP9, # DMP900, R&D Systems; Human CXCL1/GROα, # DGR00, R&D Systems; Human CXCL8/IL-8, # D8000C, R&D Systems; Human CXCL11/I-TAC, # DCX110, Human CXCL14/BRAK, # DY866, R&D Systems) following the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Control

Journal: PLoS ONE

Article Title: MMP-9 and CXCL8/IL-8 Are Potential Therapeutic Targets in Epidermolysis Bullosa Simplex

doi: 10.1371/journal.pone.0070123

Figure Lengend Snippet: A. SQRT-PCR of chemokine mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). CXCL1 , CXCL8/IL-8 and CXCL14 expression was increased in KEB-7. Only CXCL11 and CXCL14 were increased in EBDM-1. B. CXCL8/IL-8 ELISA of 48-h-conditioned cell culture supernatant showed 2-fold upregulation at the protein level in KEB-7 but not in EBDM-1, which correlates with the SQRT-PCR results (n = 4). C. CXCL8/IL-8 concentrations were highly increased in EBS patients blister fluids. In blister fluids of healthy controls no CXCL8/IL-8 was detectable (n = 3 to n = 4). The numbers correlate with . Student’s t -test was performed with p values: * ≤0.05, *** ≤0.005, Δ≤0.001, ‡ = no significant difference between investigated cell lines. (In 6C, the Student’s t -test compared the entire patient group to the entire control group).

Article Snippet: Protein levels of KLK5, MMP7, MMP9, CXCL1, CXCL8/IL-8, CXCL11 and CXCL14 were determined in 48-h-conditioned cell culture supernatant and in patients blister fluids by using Quantikine® ELISA (Human KLK5, # DKK500, R&D Systems; Human MMP7, # DMP700, R&D Systems; Human MMP9, # DMP900, R&D Systems; Human CXCL1/GROα, # DGR00, R&D Systems; Human CXCL8/IL-8, # D8000C, R&D Systems; Human CXCL11/I-TAC, # DCX110, Human CXCL14/BRAK, # DY866, R&D Systems) following the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Control

Journal: Clinical Cancer Research

Article Title: CXCL14 Promotes a Robust Brain Tumor-Associated Immune Response in Glioma

doi: 10.1158/1078-0432.ccr-21-2830

Figure Lengend Snippet: Figure 4. Increased CXCL14 expression in PXA tumors and patient-derived cell lines. A, Volcano plot showing differentially expressed genes between PXA and IDH-mt astrocytoma highlighting CXCL14 (blue box). B, Dot plot showing CXCL14 normalized mRNA count in PXA (red, n ¼ 21) versus IDH-mt (blue, n ¼ 6). C, Increased expression of CXCL14 in RAF-altered astrocytoma (n ¼ 38) versus RAF-WT astrocytoma (n ¼ 164) in a cohort of pediatric low-grade and high-grade astrocytoma. Data from CBTN downloaded via Kids First Data Resource Portal. D, Representative images from serial sections showing BRAF V600E-mutant tumor cells (brown, arrow) corresponding to regions of abundant CXCL14 mRNA-expressing cells (red, arrow). Dual immunostaining for Iba1 (green, arrowhead) highlights lack of CXCL14 expression in microglia/macrophages. E, Representative image of BRAFV600E (red), CXCL14 (yellow), and nuclei (blue) in BRAF V600E-mutant PXA. F, Expression of CXCL14 in DBTRG and patient-derived PXA lines PXA1 and PXA2 as compared with NHA-IDH1–mt cell line by qRT-PCR. G, Secreted CXCL14 protein in cell-free CM from DBTRG, PXA1, PXA2, and NHA-IDH1–mutant cell lines measured by ELISA. H, Flow cytometric analysis of cell surface HLA-ABC expression on IDH-mt after addition of CXCL14 ligand (200 ng/mL, 24 hours). In (B) and (C) each dot represents a value from a single patient; black lines represent the mean SD. In (D) red dots correspond to single RNA molecules. In F to H, data shown in box plots are mean þ SEM. Results are data from at least technical triplicates and are representative of experiments performed in biologic triplicate. Significance obtained using ANOVA and two-sided, unpaired Mann–Whitney U test. , P < 0.01; , P < 0.001; , P < 0.0001. Scale bar: D, 100 mmol/L; E, 20 mm. Ctrl, control.

Article Snippet: The CXCL14 ELISA Kit (R&D Systems; catalog no. DY866) was used according to the manufacturer’s protocol and absorbance obtained at 450 nm (Epoch Spectrophotometer, Biotech).

Techniques: Expressing, Derivative Assay, Mutagenesis, Immunostaining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Control

Journal: Clinical Cancer Research

Article Title: CXCL14 Promotes a Robust Brain Tumor-Associated Immune Response in Glioma

doi: 10.1158/1078-0432.ccr-21-2830

Figure Lengend Snippet: Figure 5. Secreted factors from tumor cells pro- mote activated CD8 T-cell chemotaxis in vitro. A, Relative chemotaxis of acti- vated CD8 T cells toward cell-free CM derived from DBTRG, PXA1, and PXA2 as compared with control media by transwell migration assay. B, Chemo- taxis of activated CD8þ T cells toward increasing concentration of CXCL14 ligand (25–100 ng/mL) by transwell migration assay. C, CXCL14 ELISA of cell-free CM after transduction of DBTRG with four pooled siRNA target- ing CXCL14 gene (siCXCL14) versus siScr. D, Chemotaxisof activatedCD8þ

Article Snippet: The CXCL14 ELISA Kit (R&D Systems; catalog no. DY866) was used according to the manufacturer’s protocol and absorbance obtained at 450 nm (Epoch Spectrophotometer, Biotech).

Techniques: Chemotaxis Assay, In Vitro, Derivative Assay, Control, Transwell Migration Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Transduction

Journal: Clinical Cancer Research

Article Title: CXCL14 Promotes a Robust Brain Tumor-Associated Immune Response in Glioma

doi: 10.1158/1078-0432.ccr-21-2830

Figure Lengend Snippet: Figure 6. Secreted CXCL14 promotes an antitumor CD8þ T-cell response in vivo. A, Secreted CXCL14 in cell-free CM from mTPCs expressing CXCL14 or GFP-control. B, Quantification of CD8þ T cells as percentage of total cells in CXCL14 (n ¼ 8 mice) and control tumors (n ¼ 5 mice). C, Representative images demonstrating CD3þ (green), CD8þ (violet), and dual CD3þCD8þ (white, arrow) T cells in CXCL14 and control tumors. D, Quantification of Iba1þ TAMs per tumor area in CXCL14 (n ¼ 7 mice) relative to control tumors (n ¼ 4 mice). E, Kaplan–Meier survival analysis. Mice with intracerebral allograft of CXCL14 mTPC have prolonged survival (median survival of 64.8 days) relative to control mTPC (median survival of 35.8 days; P < 0.003, n ¼ 8, and n ¼ 5, respectively). F, Quantification of proliferating cells as denoted by Ki-67 per tumor area in CXCL14 (n ¼ 7 mice) relative to control tumors (n ¼ 4 mice). G, Schema for tumor implantation in nude mice (n ¼ 5 per group) and FVB mice (n ¼ 10 per group) with biweekly anti-CD8a treatment to deplete CD8þ T cells (blue arrows). Corresponding isotype antibody used as control. Nude mice were implanted with either CXCL14 mTPC or control mTPC and FVB mice were implanted with CXCL14 mTPC. Kaplan–Meier survival analysis (bottom). H, Correlation between CXCL14 gene expression and CD8A gene expression in pediatric astrocytoma (CBTN; Spearman correlation coefficient, r ¼ 0.45, n ¼ 202, P < 0.0001). Bar graphs represent mean þ SEM. Significance was obtained using Student t test. Figure 6G (top FVB and nude mouse) was created with BioRender.com. , P < 0.05; , P < 0.01; , P < 0.0001; NS, P > 0.05. Scale bar: 100 mm. Ctrl, control.

Article Snippet: The CXCL14 ELISA Kit (R&D Systems; catalog no. DY866) was used according to the manufacturer’s protocol and absorbance obtained at 450 nm (Epoch Spectrophotometer, Biotech).

Techniques: In Vivo, Expressing, Control, Tumor Implantation, Gene Expression

Journal: Frontiers in Oncology

Article Title: Plasma CXCL14 as a Candidate Biomarker for the Diagnosis of Lung Cancer

doi: 10.3389/fonc.2022.833866

Figure Lengend Snippet: Plasma CXCL14 as a diagnostic biomarker in distinguishing lung cancer patients from control subjects. (A) Comparison of the CXCL14 concentration (determined by ELISA) in plasma between control subjects (n=80) and lung cancer patients (n=286) in a retrospective cohort. P <0.0001 determined by Mann–Whitney U tests. (B) ROC analysis of the diagnostic efficiency of CXCL14 in control subjects versus lung cancer patients in a retrospective cohort (AUC=0.9464, 95% CI: 0.9209–0.9719). (C) Comparison of the CXCL14 concentration (determined by ELISA) in plasma between control subjects (n=80) and stage I lung cancer patients (n=137) in a retrospective cohort. P <0.0001 determined by Mann–Whitney U tests. (D) ROC analysis of the diagnostic efficiency of CXCL14 in control subjects versus stage I lung cancer patients in a retrospective cohort (AUC=0.9353, 95% CI: 0.9034–0.9672). Scatter diagrams present the median values with interquartile ranges.

Article Snippet: The CXCL14 level in plasma and urine was assayed using commercially available sandwich enzyme-linked immunosorbent assay kits, Human CXCL14/BRAK DuoSet ELISA and DuoSet ELISA Ancillary Reagent Kit 2(Lot DY866&DY008, R&D Systems).

Techniques: Clinical Proteomics, Diagnostic Assay, Biomarker Discovery, Control, Comparison, Concentration Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Frontiers in Oncology

Article Title: Plasma CXCL14 as a Candidate Biomarker for the Diagnosis of Lung Cancer

doi: 10.3389/fonc.2022.833866

Figure Lengend Snippet: Urinary CXCL14 as a diagnostic biomarker in distinguishing lung cancer patients from control subjects. (A) Comparison of the CXCL14 concentration (determined by ELISA) in urine between control subjects (n=122) and lung cancer patients (n=284) in a retrospective cohort. P <0.0001 determined by Mann–Whitney U tests. (B) ROC analysis of the diagnostic efficiency of urinary CXCL14 in a retrospective cohort of control subjects versus lung cancer patients (AUC=0.6476, 95% CI: 0. 0.5934–0.7091, P <0.0001). Scatter diagrams present the median values with interquartile ranges. (C) Comparison of the CXCL14 concentration (determined by ELISA) in urine between control subjects (n=122) and stage I lung cancer patients (n=158) in a retrospective cohort. P <0.0001 determined by Mann–Whitney U tests. (D) ROC analysis of the diagnostic efficiency of CXCL14 in control subjects versus stage I lung cancer patients in a retrospective cohort (AUC=0.647, 95% CI: 0.5829–0.7111). Scatter diagrams present the median values with interquartile ranges.

Article Snippet: The CXCL14 level in plasma and urine was assayed using commercially available sandwich enzyme-linked immunosorbent assay kits, Human CXCL14/BRAK DuoSet ELISA and DuoSet ELISA Ancillary Reagent Kit 2(Lot DY866&DY008, R&D Systems).

Techniques: Diagnostic Assay, Biomarker Discovery, Control, Comparison, Concentration Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: The Expression of the Chemokine CXCL14 Correlates with Several Aggressive Aspects of Glioblastoma and Promotes Key Properties of Glioblastoma Cells

doi: 10.3390/ijms20102496

Figure Lengend Snippet: CXCL14 protein expression and secretion. Measurement of CXCL14 protein levels by ELISA in cell protein extracts of cultured human astrocytes, three human glioblastoma cell lines (A172, LN229, U87MG), and in protein extracts and conditioned media of NIH-ctr and NIH-CXCL14 fibroblasts. Results are shown as the mean ± S.D. and represent the average of three biological replicates per sample.

Article Snippet: The CXCL14 protein level in protein extracts and conditioned media was determined using Human CXCL14 PicoKine TM ELISA Kit (BOSTER, Pleasanton, CA, USA) following the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: The Expression of the Chemokine CXCL14 Correlates with Several Aggressive Aspects of Glioblastoma and Promotes Key Properties of Glioblastoma Cells

doi: 10.3390/ijms20102496

Figure Lengend Snippet: Exogenously supplemented CXCL14 positively affects the proliferation of glioblastoma cells. Proliferation (MTS assay) of U87MG ( A ) or LN229 ( B ) cells grown in the presence of NIH-ctr or NIH-CXCL14 conditioned media. In panel ( A ), the results of the supplementation of 10 μM AMD3100 to U87MG cells incubated with either NIH-ctr or NIH-CXCL14 conditioned media are also shown. Results are shown as the mean ± S.D. and represent the average of three experiments performed independently. Data were analyzed by a two-tailed unpaired Student’s t -test. * p < 0.05; ** p < 0.01.

Article Snippet: The CXCL14 protein level in protein extracts and conditioned media was determined using Human CXCL14 PicoKine TM ELISA Kit (BOSTER, Pleasanton, CA, USA) following the manufacturer’s protocol.

Techniques: MTS Assay, Incubation, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: The Expression of the Chemokine CXCL14 Correlates with Several Aggressive Aspects of Glioblastoma and Promotes Key Properties of Glioblastoma Cells

doi: 10.3390/ijms20102496

Figure Lengend Snippet: The treatment with CXCL14 induces the phosphorylation of ERK1 and ERK2 in U87MG cells. Representative Western blot showing total (ERK1/2) ( lower panel ) and phosphorylated (phospho-ERK1/2) ERK1/2 ( upper panel ) proteins in total protein extracts of MCF7 or U87MG cells treated or not with recombinant human CXCL14 (400 ng/mL). The graphs show the densitometric quantification of the detected bands, and represent the average (+/− st. dev.) of three independent experiments. The panel relative to MCF7 cells, assayed as positive controls of CXCL14 action on ERK phosphorylation, was produced after a longer exposure, in order to reveal the faint bands present in untreated cells. * p < 0.05.

Article Snippet: The CXCL14 protein level in protein extracts and conditioned media was determined using Human CXCL14 PicoKine TM ELISA Kit (BOSTER, Pleasanton, CA, USA) following the manufacturer’s protocol.

Techniques: Phospho-proteomics, Western Blot, Recombinant, Produced

Journal: International Journal of Molecular Sciences

Article Title: The Expression of the Chemokine CXCL14 Correlates with Several Aggressive Aspects of Glioblastoma and Promotes Key Properties of Glioblastoma Cells

doi: 10.3390/ijms20102496

Figure Lengend Snippet: Exogenously supplemented CXCL14 increases the migratory ability of LN229 and U87MG glioblastoma cells. ( A ) Representative picture (left) and relative graphical visualization (right) of a scratch test assay measuring the migration ability of LN229 cells previously incubated with either NIH-ctr or NIH-CXCL14 conditioned medium. Pictures were taken at time 0, when the scratch was performed, and after 18 h from scratching. In the graph, the distance migrated by cells in control conditions is set as = 1. ( B ) Migration transwell assays performed with LN229 cells after incubation with conditioned media of either negative control NIH-ctr or CXCL14 secreting NIH-CXCL14 cells. ( C ) Migration transwell assays performed with U87MG cells after incubation with conditioned media of either negative control NIH-ctr or CXCL14 secreting NIH-CXCL14 cells, with or without the supplementation of 10 μM AMD3100. The graph shows the number of migrating cells as compared to negative control, set as = 1. Results are presented as mean ± S.D. with significant differences from controls (*) shown ( p < 0.05). Two-tailed unpaired t -tests were used to determine significance between groups. The experiments were performed three times (biological replicates).

Article Snippet: The CXCL14 protein level in protein extracts and conditioned media was determined using Human CXCL14 PicoKine TM ELISA Kit (BOSTER, Pleasanton, CA, USA) following the manufacturer’s protocol.

Techniques: Wound Healing Assay, Migration, Incubation, Control, Negative Control, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: The Expression of the Chemokine CXCL14 Correlates with Several Aggressive Aspects of Glioblastoma and Promotes Key Properties of Glioblastoma Cells

doi: 10.3390/ijms20102496

Figure Lengend Snippet: Glioblastoma cells stably overexpressing CXCL14 show enhanced proliferation and migration. ( A ) qRT-PCR analysis of CXCL14 mRNA expression in U87MG (upper panel) or LN229 (lower panel) cells transfected with either the empty vector pcDNA3.1/V5-HisC (pcDNA), or the CXCL14-expressing vector pCXCL14-V5. In both panels, data are expressed as compared to empty vector-transfected cells, where CXCL14 expression was set as = 1. ( B ) Western blot analysis showing CXCL14 protein expression in U87MG (upper panel) or LN229 (lower panel) cells transfected with either the empty vector pcDNA3.1/V5-His C, or the CXCL14-expressing vector pCXCL14-V5. In the immunoblots shown on the left, ectopic CXCL14 was revealed by using an anti-V5 antibody, while on the right a CXCL14-specific antibody was used. β-actin detection was used as a loading control. ( C ) Proliferation (MTS assay) of U87MG (left) or LN229 (right) cells stably transfected with either the empty vector pcDNA3.1/V5-His C, or the CXCL14-expressing vector pCXCL14-V5. Results are shown as the mean ± S.D. and represent the average of two experiments performed in triplicate. Data were analyzed by a two-tailed unpaired Student’s t -test. * p < 0.05 ** p < 0.01. ( D ) Boyden chamber migration assay performed with U87MG cells stably transfected with either the empty vector pcDNA3.1/V5-HisC (pcDNA), or the CXCL14-expressing vector pCXCL14-V5. The graph depicts the relative migration of CXCL14-expressing cells compared to empty vector transfected ones, whose migration ability was set as = 1. Results are presented as mean ± S.D. with significant differences from controls (*) shown ( p < 0.05).

Article Snippet: The CXCL14 protein level in protein extracts and conditioned media was determined using Human CXCL14 PicoKine TM ELISA Kit (BOSTER, Pleasanton, CA, USA) following the manufacturer’s protocol.

Techniques: Stable Transfection, Migration, Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Western Blot, Control, MTS Assay, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: The Expression of the Chemokine CXCL14 Correlates with Several Aggressive Aspects of Glioblastoma and Promotes Key Properties of Glioblastoma Cells

doi: 10.3390/ijms20102496

Figure Lengend Snippet: Exogenously supplemented CXCL14 enhances the self-renewal ability of glioblastoma stem cells. Glioblastoma stem cell self-renewal assay. The histogram shows the fold change (FC) in numbers of neurospheres formed by three different glioblastoma stem cell lines, BT275, BT168, and BT517. The results were obtained by counting the number of neurospheres in four random fields per sample, and comparing cells grown in the presence of NIH-ctr conditioned medium to cells grown in the presence of NIH-CXCL14 conditioned medium. For BT517 cells, the assay was performed in either the presence or the absence of AMD3100 ( n = 3; mean ± S.D.). ** p -value < 0.01; *** p -value < 0.001.

Article Snippet: The CXCL14 protein level in protein extracts and conditioned media was determined using Human CXCL14 PicoKine TM ELISA Kit (BOSTER, Pleasanton, CA, USA) following the manufacturer’s protocol.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Expression of the Chemokine CXCL14 Correlates with Several Aggressive Aspects of Glioblastoma and Promotes Key Properties of Glioblastoma Cells

doi: 10.3390/ijms20102496

Figure Lengend Snippet: CXCL14 expression correlates with aggressive features in glioblastoma. ( A ) Box-plots of CXCL14 expression in the Rembrandt dataset including 315 gliomas of different grades, as analyzed by GlioVis ( http://gliovis.bioinfo.cnio.es/ ) . GBM IV versus GBM II p = 1.2E−08; GBM IV versus III p = 7.8E−09. Pairwise comparisons between group levels with corrections for multiple testing ( p -values with Bonferroni correction). ( B ) Box-plots representing the GlioVis ( http://gliovis.bioinfo.cnio.es/ ) analysis of CXCL14 expression in the IVY GAP dataset, containing RNA-seq results of a total of 122 RNA samples of five anatomic structures generated from 10 glioblastomas (as in the table below, which shows the pairwise comparisons between group levels with corrections for multiple testing ( p -values with Bonferroni correction)). ( C , D ) Box-plot graphs representing the GlioVis ( http://gliovis.bioinfo.cnio.es/ ) analysis of CXCL14 expression in the TCGA dataset, including glioblastoma samples from 528 patients, whose tumors were classified as “classical” CL, mesenchymal” MES or “proneural” PN (in C) (MES versus CL p = 3.8 × 10 −9 ; MES vs PN p = 2.7 × 10 −42 ; CL vs PN p = 1.5 × 10 −19 ) or as NON G-CIMP and G-CIMP (in D) (NON G-CIMP vs G-CIMP p = 2.8 × 10 −14 ). Pairwise comparisons between group levels with corrections for multiple testing ( p -values with Bonferroni correction). In all graphs, for each group of samples, the number in brackets represents the number of samples analyzed.

Article Snippet: The CXCL14 protein level in protein extracts and conditioned media was determined using Human CXCL14 PicoKine TM ELISA Kit (BOSTER, Pleasanton, CA, USA) following the manufacturer’s protocol.

Techniques: Expressing, RNA Sequencing, Generated

Journal: International Journal of Molecular Sciences

Article Title: The Expression of the Chemokine CXCL14 Correlates with Several Aggressive Aspects of Glioblastoma and Promotes Key Properties of Glioblastoma Cells

doi: 10.3390/ijms20102496

Figure Lengend Snippet: CXCL14 expression inversely correlates with overall survival in glioblastoma. On the left, a comprehensive graph shows the overall survival of all glioblastoma patients in the TCGA dataset, as analyzed by GlioVis ( http://gliovis.bioinfo.cnio.es/ ) ; on the right, three distinct graphs were generated by separately analyzing GBM patients based on the subtypes of their tumors.

Article Snippet: The CXCL14 protein level in protein extracts and conditioned media was determined using Human CXCL14 PicoKine TM ELISA Kit (BOSTER, Pleasanton, CA, USA) following the manufacturer’s protocol.

Techniques: Expressing, Generated

Journal: Materials Today Bio

Article Title: CXCL chemokines-mediated communication between macrophages and BMSCs on titanium surface promotes osteogenesis via the actin cytoskeleton pathway

doi: 10.1016/j.mtbio.2023.100816

Figure Lengend Snippet: Primers used for qRT-PCR.

Article Snippet: Proteins secreted by the two types of cells in the co-culture model were detected using CXCL3 and PTN ELISA Kits from JiangLai Biological (Shanghai, China) and CXCL6 and CXCL14 ELISA Kits from Cloud-Clone Corp (Wuhan, China).

Techniques: Sequencing

Journal: Materials Today Bio

Article Title: CXCL chemokines-mediated communication between macrophages and BMSCs on titanium surface promotes osteogenesis via the actin cytoskeleton pathway

doi: 10.1016/j.mtbio.2023.100816

Figure Lengend Snippet: Dynamic interactions between BMMs and BMSCs in the co-culture system. Under co-culture conditions, a reciprocal interplay unfolds between BMMs and BMSCs, driven by distinct signaling mechanisms. BMMs initiate changes in BMSCs by upregulating the secretion of chemokines and engaging chemokine receptors on BMSC surfaces. Conversely, BMSCs exert influence on BMMs by heightening PTN secretion and interacting with the Sdc3 receptors on BMM surfaces. (A) Temporal evolution of chemokine family-related gene expression in BMMs at mRNA levels in separate culture and co-culture systems. (B) Alterations in the expression of chemokine family-related gene receptors in BMSCs at the mRNA level in separate and co-culture systems. (C) Time-dependent shifts in PTN mRNA expression. (D) Temporal changes in PTN receptor Sdc3 mRNA expression on macrophages. (E, F) Protein expression profiles of CXCL3, CXCL6, CXCL14, and PTN over time in separate and co-culture systems. (G, H) Dynamics of protein expression for chemokine receptors CCR1 and CXCR2, and PTN receptor Sdc3, over time in separate and co-culture systems. (I) Time-course variations in CXCL3, CXCL6, CXCL14, and PTN levels in cell supernatant observed using ELISA in separate and co-culture systems. Data are presented as mean ± SEM of at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. mRNA and protein expression were normalized to those of the separate culture system on day 1.

Article Snippet: Proteins secreted by the two types of cells in the co-culture model were detected using CXCL3 and PTN ELISA Kits from JiangLai Biological (Shanghai, China) and CXCL6 and CXCL14 ELISA Kits from Cloud-Clone Corp (Wuhan, China).

Techniques: Co-Culture Assay, Expressing, Enzyme-linked Immunosorbent Assay