Journal: International Journal of Cell Biology

Article Title: CXCL12 Modulates Prostate Cancer Cell Adhesion by Altering the Levels or Activities of β 1-Containing Integrins

doi: 10.1155/2014/981750

Figure Lengend Snippet: Plasma CXCL12 concentrations in the 3 groups analyzed.

Article Snippet: Human recombinant CXCL12, Quantikine human CXCL12/SDF-1 immunoassay kit, and mouse anti-human CXCR4 were from R&D Systems (Minneapolis, USA).

Techniques: Clinical Proteomics, Control

Journal: International Journal of Cell Biology

Article Title: CXCL12 Modulates Prostate Cancer Cell Adhesion by Altering the Levels or Activities of β 1-Containing Integrins

doi: 10.1155/2014/981750

Figure Lengend Snippet: Mean and median plasma CXCL12 concentrations in subgroup H (Gleason score >7 including 4 + 3) were significantly higher than in subgroup L (Gleason score >7 including 4 + 3).

Article Snippet: Human recombinant CXCL12, Quantikine human CXCL12/SDF-1 immunoassay kit, and mouse anti-human CXCR4 were from R&D Systems (Minneapolis, USA).

Techniques: Clinical Proteomics

Journal: International Journal of Cell Biology

Article Title: CXCL12 Modulates Prostate Cancer Cell Adhesion by Altering the Levels or Activities of β 1-Containing Integrins

doi: 10.1155/2014/981750

Figure Lengend Snippet: (a) DU145 cell spreading on COL-I, FN, 50K, and H/120 fragment of FN in the presence of various concentrations of CXCL12 (200 ng/mL). (b) Attachment of DU145 cells on FN and COL-I in the absence and presence of CXCL12 (200 ng/mL). (c) PC3 cell spreading on COL-I, FN, 50K, and H/120 fragment of FN in the presence of various concentrations of CXCL12 (200 ng/mL). (d) PC3 cell attachment on FN and COL-I in the absence and presence of CXCL12 (200 ng/mL). The level of nonspecific binding, determined from these cells attachment to wells coated with BSA alone, was subtracted. Values shown are mean ± standard deviation of triplicate wells.

Article Snippet: Human recombinant CXCL12, Quantikine human CXCL12/SDF-1 immunoassay kit, and mouse anti-human CXCR4 were from R&D Systems (Minneapolis, USA).

Techniques: Cell Attachment Assay, Binding Assay, Standard Deviation

Journal: International Journal of Cell Biology

Article Title: CXCL12 Modulates Prostate Cancer Cell Adhesion by Altering the Levels or Activities of β 1-Containing Integrins

doi: 10.1155/2014/981750

Figure Lengend Snippet: CXCL12 induces focal adhesion disassembly, actin stress fiber rearrangement, and morphological change in DU145 cells seeded on FN. Cells were incubated for 2 hrs, fixed, and double-stained for vinculin and actin. Arrows in the images indicate localization of vinculin at the end of actin bundles in focal adhesion complexes.

Article Snippet: Human recombinant CXCL12, Quantikine human CXCL12/SDF-1 immunoassay kit, and mouse anti-human CXCR4 were from R&D Systems (Minneapolis, USA).

Techniques: Incubation, Staining

Journal: International Journal of Cell Biology

Article Title: CXCL12 Modulates Prostate Cancer Cell Adhesion by Altering the Levels or Activities of β 1-Containing Integrins

doi: 10.1155/2014/981750

Figure Lengend Snippet: CXCL12 induces focal adhesion formation, actin stress fiber rearrangement, and morphological change in PC3 cells seeded on FN. Cells were incubated for 2 hrs, fixed, and double-stained for vinculin and actin. Arrows in the images indicate localization of vinculin at the end of actin bundles in focal adhesion complexes.

Article Snippet: Human recombinant CXCL12, Quantikine human CXCL12/SDF-1 immunoassay kit, and mouse anti-human CXCR4 were from R&D Systems (Minneapolis, USA).

Techniques: Incubation, Staining

Journal: International Journal of Cell Biology

Article Title: CXCL12 Modulates Prostate Cancer Cell Adhesion by Altering the Levels or Activities of β 1-Containing Integrins

doi: 10.1155/2014/981750

Figure Lengend Snippet: PC3 cells, untreated (0) or treated with CXCL12 (200 ng/mL), were lysed and then cell lysates were analyzed by immunoblotting with anti-tyrosine-phosphorylated site-specific antibody against FAK. The total protein was detected by probing the blots with anti-FAK (bottom panel). The blot was performed three times and a representative one is shown.

Article Snippet: Human recombinant CXCL12, Quantikine human CXCL12/SDF-1 immunoassay kit, and mouse anti-human CXCR4 were from R&D Systems (Minneapolis, USA).

Techniques: Western Blot

Journal: International Journal of Cell Biology

Article Title: CXCL12 Modulates Prostate Cancer Cell Adhesion by Altering the Levels or Activities of β 1-Containing Integrins

doi: 10.1155/2014/981750

Figure Lengend Snippet: Flow cytograph analysis of α 4, β 1, and α 5 integrin subunits in untreated (C curve) and 200 ng/mL CXCL12 (T curve) treated cells. The T curves represent the binding of anti-integrin antibody on treated cells, the C curves represent the binding of anti- α 5 integrin antibody on untreated cells, and the mock curves (M) show the corresponding negative control antibody.

Article Snippet: Human recombinant CXCL12, Quantikine human CXCL12/SDF-1 immunoassay kit, and mouse anti-human CXCR4 were from R&D Systems (Minneapolis, USA).

Techniques: Binding Assay, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: Engagement of the CXCL12–CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis

doi: 10.3390/ijms23020867

Figure Lengend Snippet: Expression and release of CXCL12 by SMCs and EPCs. ( A ) Analysis of CXLC12 release using ELISA. Cell supernatants from monocultured SMCs and EPCs, treated as indicated, were collected 24 h after cultivation. * p < 0.05 vs. untreated cells; n = 6. ( B ) Real-time RT-PCR analysis of CXCL12 expression SMCs and EPCs treated as indicated. Results were normalized to CXCL12 expression in SMCs. * p < 0.05 vs. untreated cells; n = 5. ( C ) Detection of CXCL12 in MVs derived from monocultured SMCs and EPCs. Isolated MVs were lysed in RIPA buffer and CXCL12 levels were determined using ELISA. * p < 0.05 vs. non-injured SMCs, # p < 0.05 vs. SMC-MV; n = 5. ( D ) Enumeration of MVs in the supernatant of EPCs, non-injured SMCs and injured SMCs using flow cytometry with calibrated microbeads. * p < 0.05 vs. non-injured SMCs; n = 4. ( E ) Evaluation of the effect of EPC-SMC co-cultivation and engagement of CXCR4 on the release of CXCL12. Supernatants from monocultured SMCs, monocultured EPCs and EPCs co-cultured with SMCs, each in the presence or absence of a blocking CXCR4 Ab, were analyzed for CXCL12 concentration using ELISA. * p < 0.05 vs. SMCs, # p < 0.05 vs. EPC-SMC co-culture in the absence of anti-CXCR4; n = 5. ( F ) Real-time RT-PCR analysis of CXCL12 expression to test the impact of EPC-SMC co-cultivation and involvement of CXCR4. CXCL12 transcripts were determined in SMCs, EPCs and EPC-SMC co-cultures in the presence or absence of a blocking CXCR4 Ab. * p < 0.05 vs. SMCs, # p < 0.05 vs. EPC-SMC co-culture in the absence of anti-CXCR4; n = 5. ( G ) Real-time RT-PCR analysis of CXCL12 expression in SMCs treated with various doses of rCXCL12, CM-EPC or EPC-MV in the presence or absence of an anti-CXCR4 Ab. * p < 0.05 vs. untreated SMCs (control), # p < 0.05 vs. respective treatment in the absence of anti-CXCR4; n = 5. ( H ) Adhesion of EPCs to SMCs under flow conditions in vitro. EPCs pretreated with/without an anti-CXCR4 Ab were perfused in a parallel flow chamber and the number of cells EPCs adherent to the SMC monolayer was determined and expressed as adherent cells per 1 mm². For some experiments, the SMC monolayer was wounded by a linear scratch before perfusion of EPCs. * p < 0.05 vs. untreated and non-scratched SMCs (control), # p < 0.05 vs. respective treatment in the absence of anti-CXCR4; n = 4 to 6.

Article Snippet: In brief, SMCs or HUVECs rendered quiescent by serum starvation for 24 h were seeded in 96-well plates (~50% confluent cells) and subsequently treated with EPCs (5 × 10 4 ), 50 μL CM-EPC, 50 μL CM-EPC/SMC, EPC-MV, recombinant human PDGF-BB (20 ng/mL, R&D Systems) or recombinant human CXCL12 (rCXCL12; 1 to 50 ng/mL, R&D Systems) as indicated.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Derivative Assay, Isolation, Flow Cytometry, Cell Culture, Blocking Assay, Concentration Assay, Co-Culture Assay, Control, In Vitro

Journal: International Journal of Molecular Sciences

Article Title: Engagement of the CXCL12–CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis

doi: 10.3390/ijms23020867

Figure Lengend Snippet: Higher concentration of CXCL12 induce proliferation of SMCs via CXCR4. ( A ) Flow cytometry-based cell cycle analysis of SMCs treated for 24 h with EPCs in the presence or absence of anti-CXCR4 and anti-CXCL12 Abs as indicated. * p < 0.05 vs. untreated SMCs (control); n = 5. ( B – D ) Analysis of SMCs in S phase as determined 24 h after treatment as indicated. * p < 0.05 vs. untreated SMCs (control), # p < 0.05 vs. EPC treated SMCs in the absence of blocking Abs, § p < 0.05 vs. rCXCL12 50 ng/mL treated SMCs; n = 4 to 6. ( E ) Annexin V-FITC/PI staining with subsequent flow cytometry analysis to determine the rate of apoptotic SMCs treated as indicated. * p < 0.05 vs. untreated SMCs (control) for respective early and late apoptosis, # p < 0.05 vs. EPC-treated SMCs for late apoptosis; n = 5.

Article Snippet: In brief, SMCs or HUVECs rendered quiescent by serum starvation for 24 h were seeded in 96-well plates (~50% confluent cells) and subsequently treated with EPCs (5 × 10 4 ), 50 μL CM-EPC, 50 μL CM-EPC/SMC, EPC-MV, recombinant human PDGF-BB (20 ng/mL, R&D Systems) or recombinant human CXCL12 (rCXCL12; 1 to 50 ng/mL, R&D Systems) as indicated.

Techniques: Concentration Assay, Flow Cytometry, Cell Cycle Assay, Control, Blocking Assay, Staining

Journal: International Journal of Molecular Sciences

Article Title: Engagement of the CXCL12–CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis

doi: 10.3390/ijms23020867

Figure Lengend Snippet: EPCs stimulate migration of SMCs via CXCL12-CXCR4. ( A , B ) Transmigration of SMCs as analyzed in transwell chamber experiments with 8 μm pores and expressed as percentage of control. The bottom chamber contained migration medium (DMEM plus 0.5% FBS) supplemented with various doses of rCXCL12, EPCs or their secretory products in the absence or presence of blocking Abs as indicated. * p < 0.05 vs. control, # p < 0.05 vs. respective treatment in the absence of blocking Abs; n = 6. ( C , D ) SMC scratch assay. Subconfluent monolayers of SMCs, treated as indicated, were wounded linearly, and the area of the wound subsequently recovered by migrated SMCs was expressed as a percentage of the initial wound area. Representative photomicrographs ( C ) and quantified data ( D ) are shown. * p < 0.05 vs. untreated SMCs (control), # p < 0.05 vs. respective treatment in the absence of blocking Abs; n = 6.

Article Snippet: In brief, SMCs or HUVECs rendered quiescent by serum starvation for 24 h were seeded in 96-well plates (~50% confluent cells) and subsequently treated with EPCs (5 × 10 4 ), 50 μL CM-EPC, 50 μL CM-EPC/SMC, EPC-MV, recombinant human PDGF-BB (20 ng/mL, R&D Systems) or recombinant human CXCL12 (rCXCL12; 1 to 50 ng/mL, R&D Systems) as indicated.

Techniques: Migration, Transmigration Assay, Control, Blocking Assay, Wound Healing Assay

Journal: International Journal of Molecular Sciences

Article Title: Engagement of the CXCL12–CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis

doi: 10.3390/ijms23020867

Figure Lengend Snippet: Engagement of CXCL12–CXCR4 in proliferation and migration of endothelial cells. ( A ) Flow-cytometry-based cell cycle analysis of HUVECs treated for 24 h as indicated. * p < 0.05 vs. untreated HUVECs (control), # p < 0.05 vs. respective treatment in the absence of blocking Abs; n = 5. ( B ) Transmigration of HUVECs as analyzed in transwell chamber experiments and expressed as percentage of control. The bottom chamber contained migration medium (DMEM plus 0.5% FBS) supplemented with/without various doses of rCXCL12, CM-EPC or CM-EPC/SMC in the absence or presence of blocking Abs as indicated. * p < 0.05 vs. control, # p < 0.05 vs. respective treatment in the absence of blocking Abs; n = 6. ( C ) HUVEC scratch assay. Monolayers of HUVECs, treated as indicated, were wounded linearly, and the area of the wound subsequently recovered by migrated HUVECs was expressed as a percentage of the initial wound area. * p < 0.05 vs. untreated HUVECs (control), # p < 0.05 vs. respective treatment in the absence of blocking Abs; n = 6.

Article Snippet: In brief, SMCs or HUVECs rendered quiescent by serum starvation for 24 h were seeded in 96-well plates (~50% confluent cells) and subsequently treated with EPCs (5 × 10 4 ), 50 μL CM-EPC, 50 μL CM-EPC/SMC, EPC-MV, recombinant human PDGF-BB (20 ng/mL, R&D Systems) or recombinant human CXCL12 (rCXCL12; 1 to 50 ng/mL, R&D Systems) as indicated.

Techniques: Migration, Flow Cytometry, Cell Cycle Assay, Control, Blocking Assay, Transmigration Assay, Wound Healing Assay

Journal: International Journal of Molecular Sciences

Article Title: Engagement of the CXCL12–CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis

doi: 10.3390/ijms23020867

Figure Lengend Snippet: CXCL12–CXCR4-dependent control of SMC phenotype. ( A – E ) Analysis of EPC-mediated modulation of SMC phenotype and the involvement of the CXCL12–CXCR4 axis. SMCs were treated as indicated for 48 h and presence of SMA and Calponin was measured using flow cytometry. Data are expressed as mean fluorescence intensity (MFI) in % normalized to untreated SMCs (control). ( A – C ) * p < 0.05 vs. control and # p < 0.05 vs. SMCs treated with rCXCL2 50 ng/mL, ( D ) * p < 0.05 vs. SMCs co-cultured with EPCs, ( E ) * p < 0.05 vs. control and # p < 0.05 vs. the respective treatment of SMCs with CM-EPC or EPC-MV in the absence of anti-PDGFRß; n = 4 to 6. ( F ) Comparison of CXCL12 and PDGF-BB released by monocultured EPCs, monocultured SMCs and EPC-SMC co-cultures. Secreted paracrine factors present in the supernatant of the respective cultures were assessed using ELISA; n = 5.

Article Snippet: In brief, SMCs or HUVECs rendered quiescent by serum starvation for 24 h were seeded in 96-well plates (~50% confluent cells) and subsequently treated with EPCs (5 × 10 4 ), 50 μL CM-EPC, 50 μL CM-EPC/SMC, EPC-MV, recombinant human PDGF-BB (20 ng/mL, R&D Systems) or recombinant human CXCL12 (rCXCL12; 1 to 50 ng/mL, R&D Systems) as indicated.

Techniques: Control, Flow Cytometry, Fluorescence, Cell Culture, Comparison, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Engagement of the CXCL12–CXCR4 Axis in the Interaction of Endothelial Progenitor Cell and Smooth Muscle Cell to Promote Phenotype Control and Guard Vascular Homeostasis

doi: 10.3390/ijms23020867

Figure Lengend Snippet: Engagement of CXCL12–CXCR4 in EPC-mediated protection of cholesterol-induced phenotype switch. SMCs were treated as indicated for various time periods and presence of stated phenotype markers was determined using flow cytometry. Data are expressed as MFI in % normalized to untreated SMCs (control). ( A , B ) Assessment of the transformative potential of CXCL12 in the presence of SMC loading with Chol:MßCD complexes (Chol). SMCs were treated as indicated for 48 h. Anti-CXCR4 Ab and/or rCXCL12 were added to SMCs simultaneously with Chol:MßCD. * p < 0.05 vs. untreated SMCs (control), # p < 0.05 vs. SMCs treated with Chol, § p < 0.05 vs. SMCs treated with rCXCL2 50 ng/mL plus Chol:MßCD; n = 5. ( C , D , F , G , H ) Time course of SMA and CD68 expression by SMCs treated as indicated for various time periods. SMCs loaded with Chol:MßCD were either continuously co-cultured with rCXCL12 or EPCs for up to 96 h or were primary treated with Chol:MßCD for 48 h and only then subsequently exposed to rCXCL12 or EPCs for another 48 h (post-exposure); n = 4 to 6. ( E ) Analysis of CXCL12–CXCR4-dependent and EPC-mediated protection of cholesterol-induced phenotype switch after 48 h. * p < 0.05 vs. SMCs treated with Chol:MßCD, # p < 0.05 vs. SMCs co-cultured with EPCs and treated with Chol:MßCD plus EPCs (Chol.-EPC); n = 4.

Article Snippet: In brief, SMCs or HUVECs rendered quiescent by serum starvation for 24 h were seeded in 96-well plates (~50% confluent cells) and subsequently treated with EPCs (5 × 10 4 ), 50 μL CM-EPC, 50 μL CM-EPC/SMC, EPC-MV, recombinant human PDGF-BB (20 ng/mL, R&D Systems) or recombinant human CXCL12 (rCXCL12; 1 to 50 ng/mL, R&D Systems) as indicated.

Techniques: Flow Cytometry, Control, Expressing, Cell Culture

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: 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

Journal: Biochemical and biophysical research communications

Article Title: Characterization of heteromeric complexes between chemokine (C-X-C motif) receptor 4 and α 1 -adrenergic receptors utilizing intermolecular bioluminescence resonance energy transfer assays

doi: 10.1016/j.bbrc.2020.02.094

Figure Lengend Snippet: Effects of phenylephrine and CXCL12 on BRET between CXCR4 and α1a/b/d-AR. HEK293T cells were co-transfected with a fixed amount of CXCR4-Rluc and increasing amounts α1a-AR-EYFP (A), α1b-AR-EYFP (B) or α1d-AR-EYFP (C). 48 h after transfection, cells were treated with vehicle (=control), phenylephrine (PE, 200 μM) or CXCL12 (500 nM) for 5 min at 37°C before measuring BRET. Top: Representative measurements from a titration BRET experiment. Center: BRET50 from n=4 independent titration BRET experiments. Bottom: BRETmax from n=4 independent titration BRET experiments. *: p<0.05 vs. control.

Article Snippet: Reagents – Phenylephrine was purchased from Sigma-Aldrich and (C-X-C motif) chemokine ligand 12 (CXCL12) from Protein Foundry.

Techniques: Transfection, Titration