pe rat anti mouse cxcr4 antibody  (R&D Systems)

Journal: Pharmaceutics

Article Title: Homing of mRNA-Modified Endothelial Progenitor Cells to Inflamed Endothelium

doi: 10.3390/pharmaceutics14061194

Figure Lengend Snippet: Analysis of CXCR4 and PSGL-1 expressions on the surfaces of EPCs after transfection with CXCR4 and PSGL-1 mRNA. A total of 1 × 10 5 EPCs were seeded and transfected after 24 h with either 1 µg CXCR4, 1 µg PSGL-1 mRNA, or both mRNAs (mRNA cocktail). The CXCR4 and PSGL-1 expressions on the cell surfaces were analyzed 24 h post-transfection using flow cytometry. Cells treated only with medium and transfection reagent (TR) were used as negative control. Results are shown as mean + SD ( n = 3). Statistical differences were determined using one-way ANOVA multiple comparisons, followed by Bonferroni’s multiple comparisons test (** p < 0.01 and *** p < 0.001; ns: nonsignificant).

Article Snippet: Therefore, 1 × 10 5 transfected EPCs were detached, washed twice with 4% BSA/DPBS, and incubated with either PE rat anti-mouse CD162 (PSGL-1) antibody (BD Pharmingen, Heidelberg, Germany) or PE rat anti-mouse CXCR4 antibody (R&D Systems, Minneapolis, MN, USA) for 45 min at RT.

Techniques: Transfection, Flow Cytometry, Negative Control

Journal: Pharmaceutics

Article Title: Homing of mRNA-Modified Endothelial Progenitor Cells to Inflamed Endothelium

doi: 10.3390/pharmaceutics14061194

Figure Lengend Snippet: Analysis of cell viability of EPCs after transfection with CXCR4 and PSGL-1 mRNA. A total of 1 × 10 5 cells were seeded and transfected after 24 h with 1 µg of CXCR4, PSGL-1 mRNA, or both mRNAs (mRNA cocktail). The viability was analyzed 24 h post-transfection using a PrestoBlue™ cell viability assay. The viability of cells incubated only with the medium was set to 100%. Results are shown as mean + SEM ( n = 3).

Article Snippet: Therefore, 1 × 10 5 transfected EPCs were detached, washed twice with 4% BSA/DPBS, and incubated with either PE rat anti-mouse CD162 (PSGL-1) antibody (BD Pharmingen, Heidelberg, Germany) or PE rat anti-mouse CXCR4 antibody (R&D Systems, Minneapolis, MN, USA) for 45 min at RT.

Techniques: Transfection, Viability Assay, Incubation

Journal: Pharmaceutics

Article Title: Homing of mRNA-Modified Endothelial Progenitor Cells to Inflamed Endothelium

doi: 10.3390/pharmaceutics14061194

Figure Lengend Snippet: Chemotactic migration of mRNA-modified EPCs. A total of 1 × 10 5 EPCs were transfected with either 1 µg CXCR4 mRNA or with an mRNA cocktail containing 1 µg CXCR4 and 1 µg PSGL-1 mRNA. After overnight cultivation, 5 × 10 4 EPCs were seeded in 8 µm transwell inserts to analyze their migration capacity towards 50 ng/mL SDF-1α in a serum-reduced medium. Chemotactic migration was analyzed after 6 h at 37 °C by counting DAPI-stained migrated cells using ImageJ software. As controls, EPCs incubated with medium and medium containing transfection reagent (TR) were used. Scale bars represent 200 µm. Results are shown as mean + SD ( n = 4). Statistical differences were determined using one-way ANOVA multiple comparisons, followed by Bonferroni´s multiple comparisons test (**** p < 0.0001).

Article Snippet: Therefore, 1 × 10 5 transfected EPCs were detached, washed twice with 4% BSA/DPBS, and incubated with either PE rat anti-mouse CD162 (PSGL-1) antibody (BD Pharmingen, Heidelberg, Germany) or PE rat anti-mouse CXCR4 antibody (R&D Systems, Minneapolis, MN, USA) for 45 min at RT.

Techniques: Migration, Modification, Transfection, Staining, Software, Incubation

Journal: Pharmaceutics

Article Title: Homing of mRNA-Modified Endothelial Progenitor Cells to Inflamed Endothelium

doi: 10.3390/pharmaceutics14061194

Figure Lengend Snippet: Analysis of the adhesion strength of EPCs to activated endothelium using single-cell atomic force microscopy (AFM). Representative histograms of the adhesion signature of CXCR4 and PSGL-1 mRNA (mRNA cocktail) modified and native EPCs to TNF-α-activated and nonactivated endothelium and the quantification of adhesion forces are shown. Bonds formed between the cell surface ligands (PSGL-1) and molecules on HUVECs (E-selectin) broke slightly with increasing force until the cell completely detached from the HUVEC. The maximum downward force of the cantilever’s tip is referred to as detachment force (F detach ). ( n = 4). Statistical analysis was performed using two-way ANOVA, followed by Bonferroni´s multiple comparisons test (** p < 0.01 and **** p < 0.0001).

Article Snippet: Therefore, 1 × 10 5 transfected EPCs were detached, washed twice with 4% BSA/DPBS, and incubated with either PE rat anti-mouse CD162 (PSGL-1) antibody (BD Pharmingen, Heidelberg, Germany) or PE rat anti-mouse CXCR4 antibody (R&D Systems, Minneapolis, MN, USA) for 45 min at RT.

Techniques: Microscopy, Modification

Journal: Pharmaceutics

Article Title: Homing of mRNA-Modified Endothelial Progenitor Cells to Inflamed Endothelium

doi: 10.3390/pharmaceutics14061194

Figure Lengend Snippet: Dynamic adhesion of mRNA-modified EPCs to activated endothelium. ( A ) HUVECs were treated with 10 ng/mL TNF-α, and E-selectin expression was analyzed by flow cytometry to analyze the successful activation of the cells. ( n = 3). Statistical analysis was performed using a t -test (* p < 0.05 and *** p < 0.001). ( B ) EPCs were transfected with an mRNA cocktail of CXCR4 and PSGL-1 mRNA (1 µg each). After 24 h, the cells were stained with PKH26 and perfused over TNF-α-activated HUVECs in a flow chamber at 0.11 mL/min and a shear stress of 0.1 dyn/m 2 . ( B ) Representative images of rolling EPCs are shown every 2 s. Red arrows indicate a slowly rolling cell, and yellow arrows point to a fast-moving cell not interacting with the activated HUVECs. ( C ) Representative images of recordings at 1, 3, 5, and 7 min after starting the flow. ( D ) The number of EPCs adhered to HUVECs after 7 min was quantified ( n = 3). Statistical analysis was performed using two-way ANOVA, followed by Bonferroni´s multiple comparisons test (** p < 0.01).

Article Snippet: Therefore, 1 × 10 5 transfected EPCs were detached, washed twice with 4% BSA/DPBS, and incubated with either PE rat anti-mouse CD162 (PSGL-1) antibody (BD Pharmingen, Heidelberg, Germany) or PE rat anti-mouse CXCR4 antibody (R&D Systems, Minneapolis, MN, USA) for 45 min at RT.

Techniques: Modification, Expressing, Flow Cytometry, Activation Assay, Transfection, Staining, Shear

Journal: Data in Brief

Article Title: Supporting dataset of two integration-free induced pluripotent stem cell lines from related human donors

doi: 10.1016/j.dib.2021.107140

Figure Lengend Snippet: Reagents details.

Article Snippet: Differentiation Marker , CXCR4 (CD184) Mouse IgG2a PE , 1:11 , Miltenyi 130-098-354; AB_2655778.

Techniques: Immunocytochemistry, Marker, Control

Journal: Data in Brief

Article Title: Supporting dataset of two integration-free induced pluripotent stem cell lines from related human donors

doi: 10.1016/j.dib.2021.107140

Figure Lengend Snippet:

Article Snippet: Differentiation Marker , CXCR4 (CD184) Mouse IgG2a PE , 1:11 , Miltenyi 130-098-354; AB_2655778.

Techniques: Flow Cytometry, Software, Sterility, Membrane, Filtration, Expressing, In Vitro, Staining