Journal: iScience

Article Title: Oncostatin M is dispensable for the regulation of hematopoietic stem/progenitor cell traffic by neutrophils

doi: 10.1016/j.isci.2025.112646

Figure Lengend Snippet: Loss of OSM perturbs HSPC levels but not neutrophil level and circadian rhythms (A and B) Circadian levels of HSPCs (Lin - cKit + Sca1 + cells, LKS) in peripheral blood (A) and bone marrow (B) of wild-type (Wt) and Osm −/− mice at different time points. Asterisks report significance vs. ZT5 of each chart. (C and D) Osm gene expression (C) and OSM concentration in the bone marrow extracellular fluid (BMEF, D) in Wt and Osm −/− mice. (E) CXCL12 concentrations in the BMEF in Wt and Osm −/− mice. (F and G) Circadian levels of aged neutrophils in peripheral blood (F) and bone marrow (G) of Wt and Osm −/− mice at different time points. (H) Expression of selected clock genes in neutrophils and of phagocytosis genes in the BM of Wt and Osm −/− mice at Zt5 and Zt13, normalized to Ubc. Mean ± standard error is displayed in the plots. ∗ p < 0.05; ∗∗ p < 0.01; by Student’s t test (A, B, F, G, H), two-way ANOVA (C, D, E). n = 4–7 biological replicates for each strain.

Article Snippet: Commercial ELISA kits assays were used to quantify OSM (Cat. No DY495-05, Bio-Techne, Minneapolis, USA) and CXCL12 (Cat. No. MCX120, R&D Systems, Inc, Bio-Techne, Minneapolis, USA) in plasma, BMEF or culture supernatant.

Techniques: Gene Expression, Concentration Assay, Expressing

Journal: iScience

Article Title: Oncostatin M is dispensable for the regulation of hematopoietic stem/progenitor cell traffic by neutrophils

doi: 10.1016/j.isci.2025.112646

Figure Lengend Snippet: Neutrophil depletion mobilizes HSPCs (A) Depletion of peripheral blood neutrophils with anti-Ly6G antibodies in wild-type (Wt) and Osm −/− mice. (B) Depletion of bone marrow neutrophils with anti-Ly6G antibodies in Wt and Osm −/− mice. (C) HSPC (Lin - cKit + Sca1 + cells) levels in Wt and Osm −/− mice treated with anti-Ly6G antibodies or PBS control. (D) Gene expression of Cxcl12 in the bone marrow of Wt and Osm −/− mice treated with anti-Ly6G antibodies or PBS. (E) Gene expression of Cxcl12 in the bone marrow of Wt and Osm −/− mice treated with clodronate liposomes or PBS. Mean ± standard error is displayed in the plots. ∗ p < 0.05; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001 by two-way ANOVA (A-B-C-D-E).

Article Snippet: Commercial ELISA kits assays were used to quantify OSM (Cat. No DY495-05, Bio-Techne, Minneapolis, USA) and CXCL12 (Cat. No. MCX120, R&D Systems, Inc, Bio-Techne, Minneapolis, USA) in plasma, BMEF or culture supernatant.

Techniques: Control, Gene Expression, Liposomes

Journal: Biology of reproduction

Article Title: Cysteine-X-cysteine motif chemokine ligand 12 and its receptor CXCR4: expression, regulation, and possible function at the maternal-conceptus interface during early pregnancy in pigs.

doi: 10.1093/biolre/ioy147

Figure Lengend Snippet: Figure 1. Expression of CXCL12 (A) and CXCR4 (B) mRNA in the endometrium during

Article Snippet: Blots were incubated overnight at 4°C with 1 μg/ml mouse monoclonal anti-CXCL12 antibody (R&D Systems), 1 μg/ml mouse monoclonal antiCXCL12 antibody neutralized with 100 ng recombinant human CXCL12 protein (rCXCL12; R&D Systems), or 1 μg/ml isotype-matched normal mouse IgG1 (Vector Laboratories) diluted in 2% (w/v) fat-free milk in TBST.

Techniques: Expressing

Journal: Biology of reproduction

Article Title: Cysteine-X-cysteine motif chemokine ligand 12 and its receptor CXCR4: expression, regulation, and possible function at the maternal-conceptus interface during early pregnancy in pigs.

doi: 10.1093/biolre/ioy147

Figure Lengend Snippet: Figure 2. Localization of CXCL12 (A) and CXCR4 (B) proteins by

Article Snippet: Blots were incubated overnight at 4°C with 1 μg/ml mouse monoclonal anti-CXCL12 antibody (R&D Systems), 1 μg/ml mouse monoclonal antiCXCL12 antibody neutralized with 100 ng recombinant human CXCL12 protein (rCXCL12; R&D Systems), or 1 μg/ml isotype-matched normal mouse IgG1 (Vector Laboratories) diluted in 2% (w/v) fat-free milk in TBST.

Techniques:

Journal: Biology of reproduction

Article Title: Cysteine-X-cysteine motif chemokine ligand 12 and its receptor CXCR4: expression, regulation, and possible function at the maternal-conceptus interface during early pregnancy in pigs.

doi: 10.1093/biolre/ioy147

Figure Lengend Snippet: Figure 3. Immunoblot analysis of CXCL12 proteins in uterine flushings on Day 15 of

Article Snippet: Blots were incubated overnight at 4°C with 1 μg/ml mouse monoclonal anti-CXCL12 antibody (R&D Systems), 1 μg/ml mouse monoclonal antiCXCL12 antibody neutralized with 100 ng recombinant human CXCL12 protein (rCXCL12; R&D Systems), or 1 μg/ml isotype-matched normal mouse IgG1 (Vector Laboratories) diluted in 2% (w/v) fat-free milk in TBST.

Techniques: Western Blot

Journal: Biology of reproduction

Article Title: Cysteine-X-cysteine motif chemokine ligand 12 and its receptor CXCR4: expression, regulation, and possible function at the maternal-conceptus interface during early pregnancy in pigs.

doi: 10.1093/biolre/ioy147

Figure Lengend Snippet: Figure 4. Expression of CXCL12 and CXCR4 in conceptuses from Days 12 and 15 of

Article Snippet: Blots were incubated overnight at 4°C with 1 μg/ml mouse monoclonal anti-CXCL12 antibody (R&D Systems), 1 μg/ml mouse monoclonal antiCXCL12 antibody neutralized with 100 ng recombinant human CXCL12 protein (rCXCL12; R&D Systems), or 1 μg/ml isotype-matched normal mouse IgG1 (Vector Laboratories) diluted in 2% (w/v) fat-free milk in TBST.

Techniques: Expressing

Journal: Biology of reproduction

Article Title: Cysteine-X-cysteine motif chemokine ligand 12 and its receptor CXCR4: expression, regulation, and possible function at the maternal-conceptus interface during early pregnancy in pigs.

doi: 10.1093/biolre/ioy147

Figure Lengend Snippet: Figure 5. Effects of IFNG on CXCL12 and CXCR4 mRNA in endometrial explant

Article Snippet: Blots were incubated overnight at 4°C with 1 μg/ml mouse monoclonal anti-CXCL12 antibody (R&D Systems), 1 μg/ml mouse monoclonal antiCXCL12 antibody neutralized with 100 ng recombinant human CXCL12 protein (rCXCL12; R&D Systems), or 1 μg/ml isotype-matched normal mouse IgG1 (Vector Laboratories) diluted in 2% (w/v) fat-free milk in TBST.

Techniques:

Journal: Biology of reproduction

Article Title: Cysteine-X-cysteine motif chemokine ligand 12 and its receptor CXCR4: expression, regulation, and possible function at the maternal-conceptus interface during early pregnancy in pigs.

doi: 10.1093/biolre/ioy147

Figure Lengend Snippet: Figure 6. Effect of CXCL12 on pTr cell proliferation and migration. (A) RT-PCR

Article Snippet: Blots were incubated overnight at 4°C with 1 μg/ml mouse monoclonal anti-CXCL12 antibody (R&D Systems), 1 μg/ml mouse monoclonal antiCXCL12 antibody neutralized with 100 ng recombinant human CXCL12 protein (rCXCL12; R&D Systems), or 1 μg/ml isotype-matched normal mouse IgG1 (Vector Laboratories) diluted in 2% (w/v) fat-free milk in TBST.

Techniques: Migration, Reverse Transcription Polymerase Chain Reaction

Journal: Biology of reproduction

Article Title: Cysteine-X-cysteine motif chemokine ligand 12 and its receptor CXCR4: expression, regulation, and possible function at the maternal-conceptus interface during early pregnancy in pigs.

doi: 10.1093/biolre/ioy147

Figure Lengend Snippet: Figure 7. Effects of CXCL12 on migration of PBMCs and CD4+, CD8+, CD4+CD8+ T

Article Snippet: Blots were incubated overnight at 4°C with 1 μg/ml mouse monoclonal anti-CXCL12 antibody (R&D Systems), 1 μg/ml mouse monoclonal antiCXCL12 antibody neutralized with 100 ng recombinant human CXCL12 protein (rCXCL12; R&D Systems), or 1 μg/ml isotype-matched normal mouse IgG1 (Vector Laboratories) diluted in 2% (w/v) fat-free milk in TBST.

Techniques: Migration

Journal: Molecular medicine reports

Article Title: miR‑137 decreases proliferation, migration and invasion in rheumatoid arthritis fibroblast‑like synoviocytes.

doi: 10.3892/mmr.2017.8225

Figure Lengend Snippet: Figure 4. CXCL12 is a direct target of miR‑137 in RA‑FLS. (A) Predicted binding site for miR‑137 in the 3'UTR of Wt‑CXCL12; mutations in the binding sites in the Mut‑CXCL12 sequence are also indicated. (B) Relative luciferase activity of in RA‑FLS co‑transfected with either Wt‑CXCL12 or Mut‑CXCL12 3'UTR reporter plasmid and either miR‑137 mimic or miR‑NC. **P<0.01 vs. miR‑NC. The expression levels of CXCL12 (C) mRNA and (D) protein were detected in FLS isolated the RA model rat group and the normal control group by RT‑PCR and western blotting, respectively. GAPDH was used as an internal control. **P<0.01 vs. normal FLS. (E and F) RA‑FLS were transfected with either miR‑137 mimic or miR‑NC, and CXCL12 (E) mRNA and (F) protein expression levels were determined. GAPDH was used as an internal control. **P<0.01 vs. miR‑NC. CXCL12, C‑X‑C motif chemokine ligand 12; FLS, fibroblast‑like synoviocytes; miR, microRNA; Mut, mutant; NC, negative control; RA, rheumatoid arthritis; UTR, untranslated region; Wt, wild‑type.

Article Snippet: Membranes were blocked with 5% non‐fat milk in Tris‐buffered saline (TBS; Sigma‐Aldrich; Merck KGaA) for 1 h at room temperature, and incubated overnight at 4 ̊C with the following primary antibodies: Mouse monoclonal anti-human CXCL12 (1:1,000; cat no. sc‐74271) and mouse monoclonal anti‐human GAPDH (1:5,000; cat no. sc-293335); both were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Techniques: Binding Assay, Sequencing, Luciferase, Activity Assay, Plasmid Preparation, Expressing, Isolation, Control, Western Blot, Transfection, Mutagenesis, Negative Control

Journal: BMC Cancer

Article Title: COUP-TFI modifies CXCL12 and CXCR4 expression by activating EGF signaling and stimulates breast cancer cell migration

doi: 10.1186/1471-2407-14-407

Figure Lengend Snippet: Sequences of primers used in this study

Article Snippet: A goat polyclonal antibody against human CXCL12 (R&D Systems AF-310-NA), rabbit polyclonal antibody against CXCR4 (Abcam Inc. ab2074), mouse monoclonal antibody against human CXCR7/RDC1 (R&D Systems clone 11G8; MAB42273), a rabbit polyclonal antibody against COUP-TFI (Abcam Inc. ab11954) and a rabbit polyclonal antibody against HA epitope (Santa Cruz sc-805) were used for the immunofluorescence and western blot assays.

Techniques:

Journal: BMC Cancer

Article Title: COUP-TFI modifies CXCL12 and CXCR4 expression by activating EGF signaling and stimulates breast cancer cell migration

doi: 10.1186/1471-2407-14-407

Figure Lengend Snippet: COUP-TFI modifies the expression of the CXCL12 signaling axis in MCF-7 cells. (A) Characterization of the control and COUP clones. An immunofluorescence cytochemistry assay was used to detect the relative expression of HA/COUP-TFI or COUP-TFI proteins in the control (Cont.) and COUP clones. The cells were fixed and processed for immunofluorescence as described in Methods; the nuclei were stained with DAPI. (B) CXCL12 , CXCR4 , and CXCR7 mRNAs were quantified by a real-time PCR analysis from two independent MCF-7 control and COUP clones. The results were normalized to GAPDH mRNA used as an internal control. The results were expressed as the relative mRNA expression level of CXCL12 , CXCR4 , or CXCR7 . Data are the mean values ± SEM of at least three independent experiments. The asterisks indicate significant differences ( p < 0.05) between the control and COUP clones. (C) The amount of intracellular HA/COUP-TFI, COUP-TFI, CXCL12, CXCR4, and CXCR7 protein was determined from whole-cell extracts of the different MCF-7 clones and compared to total ERK. A representative western blot is shown. (D) The control and COUP clones were fixed, and an immunofluorescence cytochemistry assay was used to detect the relative expression of CXCL12, CXCR4, and CXCR7 proteins. Staining with DAPI is also presented to visualize the nucleus of the cells.

Article Snippet: A goat polyclonal antibody against human CXCL12 (R&D Systems AF-310-NA), rabbit polyclonal antibody against CXCR4 (Abcam Inc. ab2074), mouse monoclonal antibody against human CXCR7/RDC1 (R&D Systems clone 11G8; MAB42273), a rabbit polyclonal antibody against COUP-TFI (Abcam Inc. ab11954) and a rabbit polyclonal antibody against HA epitope (Santa Cruz sc-805) were used for the immunofluorescence and western blot assays.

Techniques: Expressing, Control, Clone Assay, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction, Western Blot

Journal: BMC Cancer

Article Title: COUP-TFI modifies CXCL12 and CXCR4 expression by activating EGF signaling and stimulates breast cancer cell migration

doi: 10.1186/1471-2407-14-407

Figure Lengend Snippet: COUP-TFI modulates the chromatin structure of the CXCL12 and CXCR4 gene promoters. The FAIRE assay was performed as described in Methods. Real-time PCR was performed to monitor the enrichment of DNA corresponding to the proximal promoter of the CXCL12 (A) , the CXCR4 (B) and the CXCR7 (C) genes relative to the input chromatin from the control (Cont.) and COUP clones. The data are from triplicate samples and are representative of three separate experiments. The asterisk indicates significant differences ( p < 0.05) between the control and COUP clones.

Article Snippet: A goat polyclonal antibody against human CXCL12 (R&D Systems AF-310-NA), rabbit polyclonal antibody against CXCR4 (Abcam Inc. ab2074), mouse monoclonal antibody against human CXCR7/RDC1 (R&D Systems clone 11G8; MAB42273), a rabbit polyclonal antibody against COUP-TFI (Abcam Inc. ab11954) and a rabbit polyclonal antibody against HA epitope (Santa Cruz sc-805) were used for the immunofluorescence and western blot assays.

Techniques: Real-time Polymerase Chain Reaction, Control, Clone Assay

Journal: BMC Cancer

Article Title: COUP-TFI modifies CXCL12 and CXCR4 expression by activating EGF signaling and stimulates breast cancer cell migration

doi: 10.1186/1471-2407-14-407

Figure Lengend Snippet: Estrogenic regulation of CXCL12 and CXCR4 in control and COUP clones. Control (Cont.) and COUP clones were treated with ethanol (EtOH) as the vehicle or E2 10 −8 M and ICI 10 −6 M alone or both together for 48 h. The CXCL12 (A) and CXCR4 (B) relative mRNA levels were monitored by a real-time PCR analysis, normalized to GAPDH mRNA as the internal control, and were expressed as the relative mRNA expression of CXCL12 or CXCR4 . Data are the mean ± SEM of at least three independent experiments. The asterisks indicate significant differences ( p < 0.05) between the untreated and treated control clones. The pound sign indicates significant differences ( p < 0.05) between the untreated and treated COUP clones.

Article Snippet: A goat polyclonal antibody against human CXCL12 (R&D Systems AF-310-NA), rabbit polyclonal antibody against CXCR4 (Abcam Inc. ab2074), mouse monoclonal antibody against human CXCR7/RDC1 (R&D Systems clone 11G8; MAB42273), a rabbit polyclonal antibody against COUP-TFI (Abcam Inc. ab11954) and a rabbit polyclonal antibody against HA epitope (Santa Cruz sc-805) were used for the immunofluorescence and western blot assays.

Techniques: Control, Clone Assay, Real-time Polymerase Chain Reaction, Expressing

Journal: BMC Cancer

Article Title: COUP-TFI modifies CXCL12 and CXCR4 expression by activating EGF signaling and stimulates breast cancer cell migration

doi: 10.1186/1471-2407-14-407

Figure Lengend Snippet: The effect of COUP-TFI on the CXCL12/CXCR4 axis is mediated by EGF/EGFR activation. (A) The relative expression of EGF and EGFR mRNA was monitored by a real-time PCR analysis using MCF-7 control (Cont.) and COUP clones. The results were normalized against GAPDH as the internal control and are expressed as the mean EGF or EGFR mRNA/GAPDH mRNA ratio ± SEM of at least three independent experiments. The asterisks indicate significant differences ( p < 0.05) between the control and COUP clones. (B) ERK activation was examined in the MCF-7 control (Cont.) and COUP clones after a 5- or 10-min stimulation with EGF (10 −9 M) or CXCL12 (200 ng/mL). Western blots were performed using antibodies against phospho-ERK (P-ERK) and total ERK (ERK1/2); a representative western blot is presented. The importance of EGFR-specific signaling and general ERK signaling on CXCL12 (C) and CXCR4 (D) regulation was assayed by treating the cells with EGF (10 −9 M), AG1478 (25 μM), or U0126 (25 μM) for 48 h. The CXCL12 and CXCR4 relative mRNA levels were monitored by the real-time PCR analysis, normalized to GAPDH mRNA as the internal control, and were expressed as the relative mRNA expression of CXCL12 or CXCR4 . Data are the mean ± SEM of at least three independent experiments. The asterisks indicate significant differences ( p < 0.05) between the untreated and treated control clones. The pound sign indicates significant differences ( p < 0.05) between the untreated and treated COUP clones.

Article Snippet: A goat polyclonal antibody against human CXCL12 (R&D Systems AF-310-NA), rabbit polyclonal antibody against CXCR4 (Abcam Inc. ab2074), mouse monoclonal antibody against human CXCR7/RDC1 (R&D Systems clone 11G8; MAB42273), a rabbit polyclonal antibody against COUP-TFI (Abcam Inc. ab11954) and a rabbit polyclonal antibody against HA epitope (Santa Cruz sc-805) were used for the immunofluorescence and western blot assays.

Techniques: Activation Assay, Expressing, Real-time Polymerase Chain Reaction, Control, Clone Assay, Western Blot

Journal: BMC Cancer

Article Title: COUP-TFI modifies CXCL12 and CXCR4 expression by activating EGF signaling and stimulates breast cancer cell migration

doi: 10.1186/1471-2407-14-407

Figure Lengend Snippet: COUP-TFI overexpression influences cellular responses to CXCL12. (A) The relative growth of the control (Cont.) and COUP clones was assayed with or without CXCL12 treatment for 7 days. The basal and CXCL12-induced cell growth were evaluated by MTT assays (n = 6) and determined in three independent experiments. The results are expressed as the relative cell number obtained when the control cells were treated with the vehicle control. Significant differences between the unstimulated control cells and the other conditions ( p < 0.05) are indicated with an asterisk. Significant differences between stimulated control cells and stimulated COUP clones ( p < 0.05 ) are indicated with a pound sign. (B) The migratory capacity of control (Cont.) and COUP clones was analyzed. The cells were maintained in phenol red-free DMEM/2.5% dsFBS for 48 h and then seeded in phenol red-free DMEM/0.5% dsFBS in the upper chamber of a PET 8-μm pore insert. The cells were allowed to migrate for 24 h toward the phenol red-free DMEM/2.5% dsFBS medium complemented or not with CXCL12 (200 ng/mL) and AMD3100 (50 μM). (C) CXCL12 was also added to the culture medium in the upper chamber prior to migration. The results are expressed as the mean ± SEM of the relative number of migratory cells compared to the basal migration of the control cells measured in three independent experiments. The asterisks indicate significant differences ( p < 0.05) from the basal migration of the control clones. The pound sign indicates significant differences ( p < 0.05) between two conditions linked by black lines.

Article Snippet: A goat polyclonal antibody against human CXCL12 (R&D Systems AF-310-NA), rabbit polyclonal antibody against CXCR4 (Abcam Inc. ab2074), mouse monoclonal antibody against human CXCR7/RDC1 (R&D Systems clone 11G8; MAB42273), a rabbit polyclonal antibody against COUP-TFI (Abcam Inc. ab11954) and a rabbit polyclonal antibody against HA epitope (Santa Cruz sc-805) were used for the immunofluorescence and western blot assays.

Techniques: Over Expression, Control, Clone Assay, Migration

Journal: BMC Cancer

Article Title: COUP-TFI modifies CXCL12 and CXCR4 expression by activating EGF signaling and stimulates breast cancer cell migration

doi: 10.1186/1471-2407-14-407

Figure Lengend Snippet: Box plots of CXCR4, CXCR7, CXCL12, and COUP-TFI mRNA expression in breast cancer and normal tissue. CXCR4 (A) , CXCR7 (B) , CXCL12 (C) , and COUP-TFI (D) mRNA expression was measured by real-time PCR in 23 normal breast tissue samples (NT), in 20 SBR grades 1 and 2, and in 19 SBR grades 3. The expression level was normalized by 18S RNA expression and analyzed with IQ5 software (Bio-Rad). The data are presented as whisker plots in which the horizontal bar represents the median, the grey boxes are the 25 th and 75 th percentiles, the vertical bar is the standard deviation, and the plus signs are the extreme points. All the Mann-Whitney tests were performed with Minitab 16 software, and the p value is indicated on the different graphs (ns denotes non-significant).

Article Snippet: A goat polyclonal antibody against human CXCL12 (R&D Systems AF-310-NA), rabbit polyclonal antibody against CXCR4 (Abcam Inc. ab2074), mouse monoclonal antibody against human CXCR7/RDC1 (R&D Systems clone 11G8; MAB42273), a rabbit polyclonal antibody against COUP-TFI (Abcam Inc. ab11954) and a rabbit polyclonal antibody against HA epitope (Santa Cruz sc-805) were used for the immunofluorescence and western blot assays.

Techniques: Expressing, Real-time Polymerase Chain Reaction, RNA Expression, Software, Whisker Assay, Standard Deviation, MANN-WHITNEY

Journal: Breast Cancer Research : BCR

Article Title: Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis

doi: 10.1186/bcr3053

Figure Lengend Snippet: Genes differentially regulated by IRF5 in MDA-MB-231 cells.

Article Snippet: Briefly, 100 ng/ml human recombinant CXCL12/SDF-1 (R&D Systems, Minneapolis, MN, USA) was added to 600 μl of phenol red-free DMEM medium supplemented with 10% FBS in the lower chamber.

Techniques: Expressing