Journal: Biofabrication

Article Title: A microphysiological assay for studying T-cell chemotaxis, trafficking and tumor killing.

doi: 10.1088/1758-5090/ad847f

Figure Lengend Snippet: Figure 2. 2-lane assay arrangement for T-cell chemotaxis (a) Changes made to the pre-gel additional sequence to achieve aligned collagen fibers (ECM: 2 mg ml−1 collagen mixed with 10% matrigel GFR) (b) three different assay arrangements (i), (ii) and (iii) to understand the best arrangement for robust activated CD3+ T-cell chemotaxis response to CXCL12 gradient. T-cell migration distances were analyzed and quantified at 120 h after experimental initiation (a representative image). Blue arrow indicates the direction of cell motion and the x = 0 chemotaxis distance. (c) Final assay arrangement to study immune cell chemotaxis against chemotactic stimuli. Direction of the gradient and resulting cell chemotaxis is shown. Scale bar = 1000 µm (linear mixed effects model corrected for multiple comparisons, ∗∗∗∗p-value < 0.0001).

Article Snippet: Recombinant IL2 (Cat No: 130- 097-748), CXCL12 (Cat No: 350-NS) and CXCL10 (Cat No: 266-IP) were purchased from R&D systems (Minneapolis, MN).

Techniques: Chemotaxis Assay, Sequencing, Migration

Journal: Biofabrication

Article Title: A microphysiological assay for studying T-cell chemotaxis, trafficking and tumor killing.

doi: 10.1088/1758-5090/ad847f

Figure Lengend Snippet: Figure 3. Activated CD3+ T cell responses to CXCL12 and CXCL10 gradient. Activated CD3+ T-cell chemotaxis response against (a) CXCL12 and (b) CXCL10 at 96 h within end-to-end ECM filled 2-lane plates. Numbers of T-cells under migration and T-cell migration length is shown (median T-cell migration distances indicated by black arrow). Inserts show representative images of 0 nM and 300 nM CXCL12 and CXCL10 split across two fields of images (taken at 4X magnification). (one-way ANOVA corrected for multiple comparisons, ∗p-value < 0.05, ∗∗p-value < 0.01, ∗∗∗∗p-value < 0.0001).

Article Snippet: Recombinant IL2 (Cat No: 130- 097-748), CXCL12 (Cat No: 350-NS) and CXCL10 (Cat No: 266-IP) were purchased from R&D systems (Minneapolis, MN).

Techniques: Chemotaxis Assay, Migration

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:

Article Title: Stimulation of Chondrocyte Hypertrophy by Chemokine Stromal Cell-Derived Factor 1 in the Chondro-osseous Junction during Endochondral Bone Formation

doi: 10.1016/j.ydbio.2010.02.033

Figure Lengend Snippet: To determine whether SDF-1 can diffuse into cartilage, 17-day-chicken embryonic sternal cartilage was incubated with SDF-1 (100ng/mL) or without SDF-1 for 1h, 3h, and 24h. 10 µm frozen sections were used to detect SDF-1 by immuno-fluorescent staining with mAb against SDF-1. Fluorescence microscopy showed a progressive increase in SDF-1 staining (red color) surrounding chondrocytes during the 24 h time course (A, B,C) compared to control at 24 h (D). Scale bar = 20 µm.

Article Snippet: Before collecting samples for experiment, the cells were stimulated with SDF-1 (100ng/mL; Cat# 351-FS, R&D Systems, Inc. Minneapolis, MN) for 24h or pretreated with AMD3100 for 2 h (5ug/mL; Cat# 155148–31–5, Sigma-Aldrich, St. Louis, MO), a specific inhibitor for CXCR4, before stimulation with SDF-1.

Techniques: Incubation, Staining, Fluorescence, Microscopy, Control

Journal:

Article Title: Stimulation of Chondrocyte Hypertrophy by Chemokine Stromal Cell-Derived Factor 1 in the Chondro-osseous Junction during Endochondral Bone Formation

doi: 10.1016/j.ydbio.2010.02.033

Figure Lengend Snippet: To confirm that SDF-1 induces chondrocyte hypertrophy in growth plates, 12-day-old chicken tibia growth plates were cultured in the presence of SDF-1 (100ng/mL) or in the absence of SDF-1 for 2, 4, and 6 days. 10 µm frozen sections were used to detect Type X collagen expression by immuno-fluorescent staining with mAb against type X collagen. A progressive increase in the size of the hypertrohic growth plate based on Type X collagen staining was seen. The ratio of the length of the hypertrophic zone to that of the total growth plate was calculated at the different time points (B). (* p<0.05). Scale bar = 100 µm.

Article Snippet: Before collecting samples for experiment, the cells were stimulated with SDF-1 (100ng/mL; Cat# 351-FS, R&D Systems, Inc. Minneapolis, MN) for 24h or pretreated with AMD3100 for 2 h (5ug/mL; Cat# 155148–31–5, Sigma-Aldrich, St. Louis, MO), a specific inhibitor for CXCR4, before stimulation with SDF-1.

Techniques: Cell Culture, Expressing, 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: 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: Science Advances

Article Title: Hypoxia and matrix viscoelasticity sequentially regulate endothelial progenitor cluster-based vasculogenesis

doi: 10.1126/sciadv.aau7518

Figure Lengend Snippet: ( A ) Intracardiac injection of GFP-ECFCs, followed by injection of SDF-1α loaded nonhypoxic and hypoxic hydrogels into the flank of nu/nu mice. ( B ) Twelve hours after injection, GFP-ECFCs exhibited single-cell spindle (i) or rounded (ii) morphology in nonhypoxic hydrogels. Host cells (GS-IB4 lectin) were also present as isolated, rounded cells. Under hypoxic conditions, GFP-ECFCs were present in clusters (iii and iv), with some clusters containing sprouting cells (iii). Host cells also exhibited cluster morphology under hypoxic conditions (iii and iv). ( C ) Quantification revealed a similar number of cells under both conditions and ( D ) a slight increase in percent area covered by clusters under hypoxic compared to nonhypoxic conditions. ( E ) Clusters were larger under hypoxic than under nonhypoxic conditions. ( F ) Intracardiac injection of GFP-ECFCs followed by injection of SDF-1α loaded hypoxic (ctl) and hypoxic (DPI) hydrogels into the flank of nu/nu mice. ( G ) Twelve hours after encapsulation, GFP-ECFCs were present as both single cells (ii and iii) and clusters (i and iv) under both conditions. ( H ) An increased number of GFP + cells were present in the control group, and ( I ) the percent area covered by clusters was increased in the control versus DPI-treated group. ( J ) Mean cluster sizes between the two groups were not statistically significantly different. n = 6 nu/nu mice per experiment. Graphical data in (C), (D), (H), and (I) are reported as box and whisker plots from minimum to maximum. Graphical data in (E) and (J) are reported as means ± SD, with all points denoted by dots. * P < 0.05.

Article Snippet: ICAM-1 antibody (BBA3), integrin-β2 antibody (AF1730), SDF-1α (350-NS-010), and the Human Protease Proteome Profiler Array Kit (ARY021B) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection, Isolation, Encapsulation, Control, Whisker Assay

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: Diagnostic Pathology

Article Title: LPS-induced CXCR7 expression promotes gastric Cancer proliferation and migration via the TLR4/MD-2 pathway

doi: 10.1186/s13000-019-0780-x

Figure Lengend Snippet: LPS induced changes in CXCR7 expression in SGC7901 cells. A: CXCR7 protein expression was assessed by western blotting after SGC7901 cells were stimulated for different periods of time with 500 ng/mL LPS; B: SGC7901 cells were cultured with various concentrations of LPS for 24 h, and CXCR7 protein expression was analyzed via western blotting; C: After exposure of SGC7901 cells to LPS (500 ng/ml), CXCR4 protein expression was assessed by western blotting; D and E: After pretreatment with CCX771, SGC7901 cell proliferation and migration were largely inhibited in response to CXCL12 (100 ng/ml) after 48 h of incubation with LPS. *, P < 0.05, vs. the NC group. The data are presented as the mean ± SD

Article Snippet: CXCR7 monoclonal antibody (clone MAB4227) and CXCL12 antibody (cat. no. 2716-SD-025/CF) were obtained from R&D systems, USA.

Techniques: Expressing, Western Blot, Cell Culture, Migration, Incubation