sdf 1 mab310 Search Results


cxcl12  (R&D Systems)
94
R&D Systems cxcl12
Expression and release of <t>CXCL12</t> 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.
Cxcl12, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/cxcl12/product/R&D Systems
Average 94 stars, based on 1 article reviews
cxcl12 - by Bioz Stars, 2026-05
94/100 stars
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recombinant human preadipocyte factor pref  (R&D Systems)
96
R&D Systems recombinant human preadipocyte factor pref
Expression and release of <t>CXCL12</t> 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.
Recombinant Human Preadipocyte Factor Pref, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/recombinant human preadipocyte factor pref/product/R&D Systems
Average 96 stars, based on 1 article reviews
recombinant human preadipocyte factor pref - by Bioz Stars, 2026-05
96/100 stars
  Buy from Supplier

Image Search Results


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.

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 some experiments, blocking antibodies (Abs) targeting CXCL12 (MAB310; R&D Systems), CXCR4 (MAB171; R&D Systems), CXCR7 (MAB42273; R&D Systems), PDGF-BB (AB-220; R&D Systems), the small molecule CXCR4 receptor antagonist AMD3100 (Sigma-Aldrich) or respective isotype controls were applied.

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

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.

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 some experiments, blocking antibodies (Abs) targeting CXCL12 (MAB310; R&D Systems), CXCR4 (MAB171; R&D Systems), CXCR7 (MAB42273; R&D Systems), PDGF-BB (AB-220; R&D Systems), the small molecule CXCR4 receptor antagonist AMD3100 (Sigma-Aldrich) or respective isotype controls were applied.

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

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.

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 some experiments, blocking antibodies (Abs) targeting CXCL12 (MAB310; R&D Systems), CXCR4 (MAB171; R&D Systems), CXCR7 (MAB42273; R&D Systems), PDGF-BB (AB-220; R&D Systems), the small molecule CXCR4 receptor antagonist AMD3100 (Sigma-Aldrich) or respective isotype controls were applied.

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

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.

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 some experiments, blocking antibodies (Abs) targeting CXCL12 (MAB310; R&D Systems), CXCR4 (MAB171; R&D Systems), CXCR7 (MAB42273; R&D Systems), PDGF-BB (AB-220; R&D Systems), the small molecule CXCR4 receptor antagonist AMD3100 (Sigma-Aldrich) or respective isotype controls were applied.

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

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.

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 some experiments, blocking antibodies (Abs) targeting CXCL12 (MAB310; R&D Systems), CXCR4 (MAB171; R&D Systems), CXCR7 (MAB42273; R&D Systems), PDGF-BB (AB-220; R&D Systems), the small molecule CXCR4 receptor antagonist AMD3100 (Sigma-Aldrich) or respective isotype controls were applied.

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

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.

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 some experiments, blocking antibodies (Abs) targeting CXCL12 (MAB310; R&D Systems), CXCR4 (MAB171; R&D Systems), CXCR7 (MAB42273; R&D Systems), PDGF-BB (AB-220; R&D Systems), the small molecule CXCR4 receptor antagonist AMD3100 (Sigma-Aldrich) or respective isotype controls were applied.

Techniques: Flow Cytometry, Control, Expressing, Cell Culture