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: bioRxiv

Article Title: Identification of CD317-Positive Pro-inflammatory Immune Stromal Cells in Human Mesenchymal Stromal Cell Preparations

doi: 10.1101/2022.02.10.479972

Figure Lengend Snippet: (A) Comparative mRNA expression of ICAM-1 in MSC lines and primary cells sorted by CD317 expression (RNA was extracted from 3 different donors or 5 cell line passages; qPCR performed in triplicate, mean shown ± SEM). (B) Mean fluorescence intensity of ICAM-1 expression on the cell surface of MSC lines and primary MSCs differentially gated by CD317 staining (MSCs from 5 different donors or 4 different passages of MSC lines were stained for flow cytometry, mean shown ± SEM). (C)/(D) Comparative (mean ± SEM) mRNA expression of CXCL10 (red) and CXCL11 (blue) in MSC lines/ primary MSCs sorted for CD317 expression (RNA was extracted from 7 different donors/7 different cell passages; experiments were performed in triplicate). (E/F) CXCL10 secretion by MSC lines prior to IFN-γ priming and after priming with baseline (unprimed) secretion subtracted (mean ± SEM, n=2). (G/H) Comparative mRNA expression of 8 IFN-γ signature genes in MSC lines/primary MSCs sorted by CD317 expression (RNA was extracted from 5 different donors/5 different cell passages; experiments were performed in triplicate, mean shown ± SEM). (I)/(J) IFN-γ score for MSC lines/primary MSCs sorted by CD317 expression (n=5)*/** = significance at P<0.05/0.01 using an appropriate statistical test.

Article Snippet: To detect secreted proteins, supernatants from 100,000 cells incubated in 2.5 ml of serum free DMEM for 24 hours was analysed for secreted proteins by enzyme-linked immunosorbent assays (ELISA) using ELISA kits for CXCL10, CXCL11 (BioLegend); CCL2 (eBioscience); and SAA4 (Stratech) following manufacturers instructions.

Techniques: Expressing, Fluorescence, Staining, Flow Cytometry

Journal: Indian Journal of Dermatology

Article Title: The Effect of Transplantation of Cultured Autologous Melanocytes on CXCL9, CXCL10 and CXCL11 Expressions in Vitiligo

doi: 10.4103/ijd.ijd_925_22

Figure Lengend Snippet: Expression levels of serum CXCL9, CXCL10 and CXCL11 in different types of vitiligo patients

Article Snippet: Before the intervention, 5 ml of peripheral venous blood (PBMC) was collected from all the participants to detect the concentrations of CXCL9, CXCL10 and CXCL11 by ELISA kit (Wuhan, Boster Biological Technology, China).

Techniques: Expressing

Journal: Indian Journal of Dermatology

Article Title: The Effect of Transplantation of Cultured Autologous Melanocytes on CXCL9, CXCL10 and CXCL11 Expressions in Vitiligo

doi: 10.4103/ijd.ijd_925_22

Figure Lengend Snippet: Expression levels of serum CXCL9, CXCL10 and CXCL11 in vitiligo patients and healthy controls

Article Snippet: Before the intervention, 5 ml of peripheral venous blood (PBMC) was collected from all the participants to detect the concentrations of CXCL9, CXCL10 and CXCL11 by ELISA kit (Wuhan, Boster Biological Technology, China).

Techniques: Expressing

Journal: Indian Journal of Dermatology

Article Title: The Effect of Transplantation of Cultured Autologous Melanocytes on CXCL9, CXCL10 and CXCL11 Expressions in Vitiligo

doi: 10.4103/ijd.ijd_925_22

Figure Lengend Snippet: Expressions levels of serum CXCL9, CXCL10 and CXCL11 in vitiligo patients of different stages

Article Snippet: Before the intervention, 5 ml of peripheral venous blood (PBMC) was collected from all the participants to detect the concentrations of CXCL9, CXCL10 and CXCL11 by ELISA kit (Wuhan, Boster Biological Technology, China).

Techniques:

Journal: Indian Journal of Dermatology

Article Title: The Effect of Transplantation of Cultured Autologous Melanocytes on CXCL9, CXCL10 and CXCL11 Expressions in Vitiligo

doi: 10.4103/ijd.ijd_925_22

Figure Lengend Snippet: Expression levels of serum CXCL9, CXCL10 and CXCL11 in stable vitiligo before and after the operation

Article Snippet: Before the intervention, 5 ml of peripheral venous blood (PBMC) was collected from all the participants to detect the concentrations of CXCL9, CXCL10 and CXCL11 by ELISA kit (Wuhan, Boster Biological Technology, China).

Techniques: Expressing

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A. Cxcl9, Cxcl10 and Cxcl11 transcript levels were measured in islets isolated from 10 week old male (n = 4) and female NOD mice (n = 5). Data are expressed relative to age-matched BALB/c control mice (n = 7). **p<0.01, *p<0.05. B–F. Islets from Wistar rats (B, C; n = 4 per group) or human islets (D–F; n =3 per group) were untreated (NT) or stimulated with 10 ng/mL IL-1β, 100 U/mL IFN-γ or both cytokines for 3 h. B–F. Relative mRNA abundance of CXCL9 (B, D), CXCL10 (E) and CXCL11 (C, F) was determined by RT-PCR. ***p<0.001 vs. NT, **p<0.01 vs. NT, *p<0.05 vs. NT.

Article Snippet: ELISA Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Isolation, Control, Reverse Transcription Polymerase Chain Reaction

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A–C. 832/13 rat insulinoma cells were stimulated with 1 ng/mL IL-1β or IL-1β plus 100 U/mL IFN-γ for the indicated times (NT; no treatment). D–F. 832/13 cells were pretreated for 1 h with either DMSO or 0.5 μg/mL Cycloheximide (CHX). Cells were subsequently exposed to IL-1β (1 ng/mL) or the combination of IL-1β and IFN-γ (100 U/mL) for 2 h. Cellular mRNA levels of Cxcl9 (A, D), Cxcl10 (B, E) and Cxcl11 (C, F) were detected by RT-PCR. n.s. = not significant vs respective treatment in DMSO control group. Data are shown as means ± SEM from three independent experiments.

Article Snippet: ELISA Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Reverse Transcription Polymerase Chain Reaction, Control

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A. 832/13 cells were exposed to 100 U/mL IFN-γ for the indicated times. PO4-STAT1Y701 and total STAT1 protein abundance were determined by immunoblotting. B, C. 832/13 cells were pre-treated for 1 h with increasing concentrations of JAKi (1 nM, 10 nM, 100 nM), followed by a 3 h stimulation with IL-1β alone (1 ng/mL) or IL-1β plus 100 U/mL IFN-γ. ***p<0.001 vs. DMSO (black bar), *p<0.05 vs. DMSO (black bar). D, E. 832/13 cells were transfected with two siRNA duplexes targeting STAT1 using a scrambled siRNA sequence duplex as a control. 48 h post- transfection cells were cultured for 3 h with 1 ng/ml IL-1β or IL-1β plus 100 U/ml IFN-γ. ***p<0.001 vs. siScramble (black bar), *p<0.05 vs. siScramble (black bar). Cxcl9 (B, D) and Cxcl11 (C, E) mRNA levels were quantified. Data are represented as means ± SEM from three independent experiments. The immunoblot in A was repeated on two separate occasions.

Article Snippet: ELISA Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Quantitative Proteomics, Western Blot, Transfection, Sequencing, Control, Cell Culture

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A. 832/13 cells were transduced with adenoviruses encoding either βGAL, wild-type STAT1 (WT), STAT1Y701F, STAT1S727A, STAT1Y701F/S727A (DM; double mutant) or STAT1S727T. STAT1 abundance was determined by immunoblotting. B, C. 832/13 cells were transduced with the adenoviruses indicated in (A); 24 h post-transduction cells were stimulated for 3 h with either IL-1β (1 ng/mL) alone or IL-1β plus IFN-γ (100 U/mL). *p<0.05, #p<0.1. D, E. Rat islets were transduced with the indicated adenoviruses. 24 h post-transduction cells were stimulated with both IL-1β (10 ng/mL) and IFN-γ (100 U/mL) for 3 h. ***p<0.001, **p<0.01. Relative mRNA abundance of Cxcl9 (B, D) and Cxcl11 (C, E) was determined by RT-PCR. Date are expressed as means ± SEM from 3 (B, C) or 2 (D, E) individual experiments. The immunoblot in A was repeated on two individual occasions.

Article Snippet: ELISA Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Transduction, Mutagenesis, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: Schematic representation of distal and proximal GAS sites in the Cxcl9 and Cxcl11 promoters are shown (left panels). Arrows are the diagrammatic depiction of PCR amplicons. A–D. 832/13 cells were stimulated with 100 U/mL IFN-γ for either 20 mins (middle panels) or a time course (right panels). ChIP assays were performed to determine relative occupancy of total STAT1 (middle panels) and PO4-STAT1Y701 (right panels) on the Cxcl9 proximal (A) and distal promoter (B), and on the Cxcl11 proximal (C) and distal (D) promoter. ***p<0.001 vs. NT, **p<0.01 vs. NT, *p<0.05 vs. NT. Data are expressed as means ± SEM from 3–4 individual experiments.

Article Snippet: ELISA Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques:

Journal: Translational Oncology

Article Title: Oncosuppressive role of MicroRNA-205–3p in gastric cancer through inhibition of proliferation and induction of senescence

doi: 10.1016/j.tranon.2021.101199

Figure Lengend Snippet: Down-regulated expression of miR-205–3p and up-regulated expression of CXCL11 in GC cells A: miRNA quantitative RT-PCR analysis of miR-205–3p expression level in eight GC cell lines and one normal gastric cell line. B: Venn diagram of the numbers of overlapping miRNAs from different databases (Targetscan, miRDB, RNA22 and DIANA). C: The miR-205–3p seed sequence is complementary to the 3′-UTR of CXCL11 and is conserved in six different species. D: Quantitative RT-PCR analysis of CXCL11 expression level in eight GC cell lines and one normal gastric cell line. E: CXCL11 expression was analyzed by Western blot in eight GC cell lines and one normal gastric cell line. * p <0.05 and ** p <0.01.

Article Snippet: The antibodies used were: anti-CXCL11 (ab216157, Abcam), anti-p-AKT (ab38449, Abcam), anti-AKT (ab8805, Abcam), anti-PD-L1 (ab205921, Abcam), anti-P16INK4A (ab201980, Abcam), anti-P21 (ab109520, Abcam), and anti-GAPDH (ab8245, Abcam).

Techniques: Expressing, Quantitative RT-PCR, Sequencing, Western Blot

Journal: Translational Oncology

Article Title: Oncosuppressive role of MicroRNA-205–3p in gastric cancer through inhibition of proliferation and induction of senescence

doi: 10.1016/j.tranon.2021.101199

Figure Lengend Snippet: miR-205–3p inhibits proliferation and invasion and promotes apoptosis by regulating CXCL11 in gastric cancer cells. A: Putative miR-205–3p binding sequence on CXCL11 wild-type(WT) 3′-UTR and mutant 3′-UTR sequence that abolished binding. B: Luciferase assay shows decreases in reporter activity after co-transfection of CXCL11 wild-type(WT) 3′-UTR with miR-205–3p mimics in AGS cells. C: In CCK8 experiment, the growth rate of AGS cells after miR-205–3p transfection is reduced compared with the NC group(miR-NC), and significantly restores after adding CXCL11(100 ng/ml). D: miR-205–3p overexpression significantly inhibits the clone-forming ability of gastric cancer cells, and the inhibition is counteracted by adding CXCL11(100 ng/ml). E: Apoptotic analysis is performed on miR-205, miR-205+CXCL11 and NC groups. The percentage of apoptotic cells is reported. F: In Transwell invasion assay, miR-205–3p overexpression significantly reduces the number of invasive cells, and the reduction is counteracted by adding CXCL11(100 ng/ml). * p <0.05 and ** p <0.01.

Article Snippet: The antibodies used were: anti-CXCL11 (ab216157, Abcam), anti-p-AKT (ab38449, Abcam), anti-AKT (ab8805, Abcam), anti-PD-L1 (ab205921, Abcam), anti-P16INK4A (ab201980, Abcam), anti-P21 (ab109520, Abcam), and anti-GAPDH (ab8245, Abcam).

Techniques: Binding Assay, Sequencing, Mutagenesis, Luciferase, Activity Assay, Cotransfection, Transfection, Over Expression, Inhibition, Transwell Invasion Assay

Journal: Translational Oncology

Article Title: Oncosuppressive role of MicroRNA-205–3p in gastric cancer through inhibition of proliferation and induction of senescence

doi: 10.1016/j.tranon.2021.101199

Figure Lengend Snippet: Effects of miR-205–3p on cell cycle progression and cell senescence. A, B: Cell cycle analysis shows increase in the G1 phase of AGS cells overexpressing miR-205 as compared with negative control (miR-NC), while additional CXCL11(100 ng/ml) reverses the increase. C, D: SA-β-gal staining was performed on miR-205, miR-205+CXCL11 and NC groups and light microscope cell images were acquired. The percentage of SA-β-gal-positive cells is reported (D). ** p <0.01.

Article Snippet: The antibodies used were: anti-CXCL11 (ab216157, Abcam), anti-p-AKT (ab38449, Abcam), anti-AKT (ab8805, Abcam), anti-PD-L1 (ab205921, Abcam), anti-P16INK4A (ab201980, Abcam), anti-P21 (ab109520, Abcam), and anti-GAPDH (ab8245, Abcam).

Techniques: Cell Cycle Assay, Negative Control, Staining, Light Microscopy

Journal: Translational Oncology

Article Title: Oncosuppressive role of MicroRNA-205–3p in gastric cancer through inhibition of proliferation and induction of senescence

doi: 10.1016/j.tranon.2021.101199

Figure Lengend Snippet: miR-205–3p inducing senescence of gastric cancer cells and secreting SASP factors by regulating CXCL11 and inhibiting Akt activation A, B: AGS cells are transfected with miR-205 alone for 48 h or with addition of CXCL11 (100 ng/mL) after 24 h of transfection for another 24 h, or transfected with CXCL11 siRNA for 48 h. and cell lysates were collected for Western blot analysis. C, D: AGS cells are treated with LY294002 (an Akt inhibitor, 25 μM) for 2 h or transfected with miR-205 for 48 h, The protein levels of indicated genes were measured by Western blot. GAPDH was used as an internal control for the total protein measurement. E: mRNA levels of the indicated genes in AGS cells transfected with negative control or miR-205. Actin was used for normalizing the expression of mRNA. * P <0.05 and ** P < 0.01.

Article Snippet: The antibodies used were: anti-CXCL11 (ab216157, Abcam), anti-p-AKT (ab38449, Abcam), anti-AKT (ab8805, Abcam), anti-PD-L1 (ab205921, Abcam), anti-P16INK4A (ab201980, Abcam), anti-P21 (ab109520, Abcam), and anti-GAPDH (ab8245, Abcam).

Techniques: Activation Assay, Transfection, Western Blot, Negative Control, Expressing