Journal: Advanced Science

Article Title: Integrated Single‐Cell and Spatial Analysis Reveals a Metabolic‐Immune Axis Driving Aortic Dissection

doi: 10.1002/advs.75509

Figure Lengend Snippet: Lentivirus‐mediated Eno1 knockdown attenuates disease progression in AAD mouse models. (A) Images showing the characteristics of aortas from four different mouse model groups: Normal (control group), AAD (BAPN+Ang‐II‐induced AAD model), AAD‐lv‐sh‐NC (empty vector transfection+Ang‐II‐induced AAD model), and AAD‐lv‐sh‐ Eno1 ( Eno1 knockdown mouse model of AAD induced by BAPN and Ang‐II). (B) Maximal abdominal aortic outer diameters in the four groups. (C) Incidence of AAD in the four groups. n = 10 per group. (D) Survival curves for mice in the four different mouse model groups. (E) H&E staining of aortic tissues from the four different model groups. (F,G) Western blot analysis of key proteins involved in contractile SMC (α‐SMA), synthetic SMC (Opn, Mmp9), and SMC_C1_CP+ (Ceruloplasmin, Vegfa) in aortic tissues from the four different model groups. (H) Immunohistochemistry showing synthetic smooth muscle marker Opn and SMC_C1_CP+ markers (Ceruloplasmin, Vegfa) in aortic tissues from the four different model groups. (I) Western blot analysis of Mif and its receptors in aortic tissues from the four different model groups. (J) Immunofluorescence showing Mif and Cxcr4 expression in aortic tissues from the four different model groups (Mif: green, Cxcr4: red). Scale bar = 50 µm. (K) Expression of anti‐inflammatory ( iNos , Cd38 ) and pro‐inflammatory factors ( Cd206 ) in aortic tissues from the four different model groups. (L) Expression of pro‐inflammatory ( Tnf‐α , Il‐6 , Il‐1β ) and anti‐inflammatory markers ( Il‐10 ) from the four different model groups. (M) Flow cytometry analysis of the ratio of Cd206+Cd86‐ M2 macrophages and Cd86+Cd206− M1 macrophages in aortic tissues from the four different model groups. For panels B, G, I, K, L and M, ANOVA followed by Tukey's post hoc test was performed for comparisons among multiple groups. *** p < 0.001; ## p < 0.01.

Article Snippet: In some experiments, THP‐1 macrophages were treated with 2 μg/mL CXCR4 antagonist AMD3100 (HY‐10046, MedChem Express, Monmouth Junction, USA), 10 ng/ml recombinant human MIF protein (r MIF ; 10246‐H08H, Sino Biological, Beijing, China), or vehicles for 24 h before functional evaluations.

Techniques: Knockdown, Biomarker Discovery, Control, Plasmid Preparation, Transfection, Staining, Western Blot, Immunohistochemistry, Marker, Immunofluorescence, Expressing, Flow Cytometry

Journal: Bioscience Reports

Article Title: Biological and mutational analyses of CXCR4–antagonist interactions and design of new antagonistic analogs

doi: 10.1042/BSR20230981

Figure Lengend Snippet: Sequences and structures of eleven antagonists

Article Snippet: AMD11070 was obtained from MedChemExpress (Monmouth Junction, NJ, U.S.A.).

Techniques: Sequencing

Journal: Bioscience Reports

Article Title: Biological and mutational analyses of CXCR4–antagonist interactions and design of new antagonistic analogs

doi: 10.1042/BSR20230981

Figure Lengend Snippet: Competitive receptor binding activities of various antagonists for wild-type CXCR4 and its mutants ^*

Article Snippet: AMD11070 was obtained from MedChemExpress (Monmouth Junction, NJ, U.S.A.).

Techniques: Binding Assay

Journal: Bioscience Reports

Article Title: Biological and mutational analyses of CXCR4–antagonist interactions and design of new antagonistic analogs

doi: 10.1042/BSR20230981

Figure Lengend Snippet: The residues implicated in antagonist binding as determined from mutational experiments are shown for ( A ) CVX15, ( B ) LY2510924, ( E ) IT1t, ( F ) AMD3100, and ( I ) AMD11070. The residues are colored in the same way and meaning as described above in . Molecular docking calculations were performed for LY2510924, AMD3100 and AMD11070 to predict the CXCR4-antagonist complex structures. Using the co-crystal structure of CXCR4-CVX15 as a reference, the superposition of each of the above-mentioned antagonists (shown in blue) with CVX15 (red) is shown for ( C ) LY2510924, ( G ) AMD3100, and ( J ) AMD11070. The interactions of LY2510924, AMD3100, and AMD11070 with CXCR4 residues based on the docking results are illustrated for ( D ) LY2510924, ( H ) AMD3100, and ( K ) AMD11070. The molecular docking calculation was conducted using the crystal structure 3OE0 and the SYBYL program and displayed by PyMOL. The CXCR4 structure (in pale cyan) is shown in ribbons with important residues highlighted in colors: D262 (yellow), E288 (Violet), W94 (Magenta), D171 (Green), and D97 (Orange). The results were averages of at least three independent experiments.

Article Snippet: AMD11070 was obtained from MedChemExpress (Monmouth Junction, NJ, U.S.A.).

Techniques: Binding Assay

Journal: Bioscience Reports

Article Title: Biological and mutational analyses of CXCR4–antagonist interactions and design of new antagonistic analogs

doi: 10.1042/BSR20230981

Figure Lengend Snippet: Binding areas of the antagonists are shown by circles or ovals in different colors of deep blue for HC4319, pink for DV1, red for DV1 dimer, green for DV3, yellow for V1, magenta for vMIP-II, cyan for CVX15, black for LY2510924, orange for IT1t, gray for AMD3100, and deep sky blue for AMD11070. The CXCR4 helices in Figures 4–7 were generated and downloaded from GPCRDB.

Article Snippet: AMD11070 was obtained from MedChemExpress (Monmouth Junction, NJ, U.S.A.).

Techniques: Binding Assay, Generated

Journal: PLoS ONE

Article Title: Comparison of cell-based assays for the identification and evaluation of competitive CXCR4 inhibitors

doi: 10.1371/journal.pone.0176057

Figure Lengend Snippet: (A), (B) and (C): Inhibition of the CXCL12/CXCR4 interaction by (A) the peptide analogs T22, T140, TC14012 and CTCE-9908 and the small molecules (B) AMD3100, AMD3465, AMD11070, IT1t and (C) WZ811, Me6TREN and gambogic acid. Jurkat cells were incubated with 2.9 nM CXCL12 AF647 in the presence of different concentrations of compound. Data are represented as % inhibition of maximal CXCL12 binding to CXCR4. Mean ± SEM of three or four independent experiments is shown.

Article Snippet: The compounds TC14012 (MW: 2,066.4 g/mol) [ ], CTCE-9908 (MW: 1,927.3 g/mol) [ ], IT1t (MW: 479.6 g/mol) [ ] and AMD3465 Hexahydrobromide (MW: 896.1 g/mol) [ ] were obtained from Tocris (Bristol, UK).

Techniques: Inhibition, Incubation, Binding Assay

Journal: PLoS ONE

Article Title: Comparison of cell-based assays for the identification and evaluation of competitive CXCR4 inhibitors

doi: 10.1371/journal.pone.0176057

Figure Lengend Snippet: Fluo-2 AM loaded cells were treated with serial dilutions of compound and subsequently stimulated with 6.25 nM CXCL12. Fluorescence changes were measured over time in all wells simultaneously. (A): The inhibitory effect of different concentrations of AMD11070 on CXCL12-induced calcium release is shown. The red and blue line represent the negative (untreated, without CXCL12 stimulation) and positive (untreated, CXCL12 stimulation) control, respectively. Calcium fluxes are represented as % of baseline ( i . e . mean relative light units in each well during a fixed time interval before CXCL12 addition). One representative experiment out of three is shown with each data point corresponding to the mean fluorescence of three to six replicates. (B), (C) and (D): Inhibitory effect of (B) T22, T140, TC14012, CTCE-9908, (C) AMD3100, AMD3465, AMD11070, IT1t, (D) WZ811, Me6TREN and gambogic acid on CXCL12-induced calcium flux. Data are represented as % inhibition of CXCL12-induced calcium mobilization relative to the positive and negative control. Mean ± SEM of three independent experiments is shown.

Article Snippet: The compounds TC14012 (MW: 2,066.4 g/mol) [ ], CTCE-9908 (MW: 1,927.3 g/mol) [ ], IT1t (MW: 479.6 g/mol) [ ] and AMD3465 Hexahydrobromide (MW: 896.1 g/mol) [ ] were obtained from Tocris (Bristol, UK).

Techniques: Fluorescence, Control, Inhibition, Negative Control

Journal: PLoS ONE

Article Title: Comparison of cell-based assays for the identification and evaluation of competitive CXCR4 inhibitors

doi: 10.1371/journal.pone.0176057

Figure Lengend Snippet: SUP-T1 cells were stained with a mAb targeting the N-terminus of CXCR4 (clone 1D9) and then treated with different concentrations of compound. CXCR4 internalization was induced by stimulating samples with CXCL12 at 37°C and quantified by flow cytometry. (A): Flow cytometric analysis of the negative (untreated, no CXCL12 stimulation; red histogram) and positive (untreated, CXCL12 stimulation; dark blue histogram) control and TC14012-treated samples (50 nM to 0.5 nM; light blue, green and orange histogram, respectively). (B), (C) and (D): Inhibitory effect of (B) T22, T140, TC14012, CTCE-9908, (C) AMD3100, AMD3465, AMD11070, IT1t, (D) WZ811, Me6TREN and gambogic acid on CXCR4 endocytosis. Data are represented as % of CXCR4 internalization relative to the positive and negative control. Mean ± SEM of at least two independent experiments is shown.

Article Snippet: The compounds TC14012 (MW: 2,066.4 g/mol) [ ], CTCE-9908 (MW: 1,927.3 g/mol) [ ], IT1t (MW: 479.6 g/mol) [ ] and AMD3465 Hexahydrobromide (MW: 896.1 g/mol) [ ] were obtained from Tocris (Bristol, UK).

Techniques: Staining, Flow Cytometry, Control, Negative Control

Journal: PLoS ONE

Article Title: Comparison of cell-based assays for the identification and evaluation of competitive CXCR4 inhibitors

doi: 10.1371/journal.pone.0176057

Figure Lengend Snippet: (A), (B) and (C): Effect of (A) T22, T140, TC14012, CTCE-9908, (B) AMD3100, AMD3465, AMD11070, IT1t, (C) WZ811, Me6TREN and gambogic acid on chemotaxis of CXCR4-positive Jurkat cells. After compound treatment, cells were allowed to migrate towards 6.25 nM CXCL12, present in the lower compartment of the insert, for two hours. The migrated cells were counted by flow cytometry. Data are represented as % inhibition of migration relative to the negative and positive control (spontaneously migrated cells and cells migrated towards CXCL12, respectively). ± SEM of two to three independent experiments, each performed in triplicate.

Article Snippet: The compounds TC14012 (MW: 2,066.4 g/mol) [ ], CTCE-9908 (MW: 1,927.3 g/mol) [ ], IT1t (MW: 479.6 g/mol) [ ] and AMD3465 Hexahydrobromide (MW: 896.1 g/mol) [ ] were obtained from Tocris (Bristol, UK).

Techniques: Chemotaxis Assay, Flow Cytometry, Inhibition, Migration, Positive Control

Journal: PLoS ONE

Article Title: Comparison of cell-based assays for the identification and evaluation of competitive CXCR4 inhibitors

doi: 10.1371/journal.pone.0176057

Figure Lengend Snippet: Anti-HIV-1 (X4) activity of CXCR4 compounds evaluated in CD4-positive MT-4 cells and PHA-stimulated PBMCs.

Article Snippet: The compounds TC14012 (MW: 2,066.4 g/mol) [ ], CTCE-9908 (MW: 1,927.3 g/mol) [ ], IT1t (MW: 479.6 g/mol) [ ] and AMD3465 Hexahydrobromide (MW: 896.1 g/mol) [ ] were obtained from Tocris (Bristol, UK).

Techniques: Activity Assay

Journal: OALib

Article Title: The Complementary Roles of CXCR4 and CXCR7 in Melanoma Migration

doi: 10.4236/oalib.1112617

Figure Lengend Snippet: Figure 1. AMD070 and ACT-1004 reduce B16-F10 migration in wound assays. B16-F10 melanoma cells were plated in 6-well plates along with either a 30 µM concentration of their respective inhibitor or 20 µL of DMSO and left in a 37˚C incubator for 24. After 24 hours, the cells were scratch wounded with a 200 µL pipette tip, and images were taken of wounds at 0, 4, 8, and 18 hours after wounding. The photos taken at 18 hours post-wounding show the wound had healed entirely in our control cells, while our control + DMSO cells were about 80% healed. The CXCR4 inhibitor (AMD070) and CXCR7 inhibitor (ACT-1004) still showed a large wound.

Article Snippet: CXCR Inhibition The CXCR4 inhibitor AMD070 hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) and the CXCR7 inhibitor ACT-1004-1239 (MedChemExpress, Monmouth Junction, NJ, USA) were diluted to 2 mg/mL and 5 mg/mL with dimethyl sulfoxide (DMSO), respectively, according to manufacturer protocols.

Techniques: Migration, Concentration Assay, Transferring, Control

Journal: OALib

Article Title: The Complementary Roles of CXCR4 and CXCR7 in Melanoma Migration

doi: 10.4236/oalib.1112617

Figure Lengend Snippet: Figure 2. CXCR inhibitors decreased melanoma wound healing by ~60%. Bar graphs showing progressive healing of melanoma wounds at each time point over an 18-hour time frame after CXCR inhibition. The data starts at 0% of the wound area healed at 0 hr, and as time progresses, cells invade the wound and make the area smaller, healing the wound. A difference was observed between the control and the control with DMSO, but an even more significant difference was observed between our controls and our AMD070 and ACT-1004 inhibited cells. *p < 0.05, **p < 0.005, ****p < 0.0001. N = 4.

Article Snippet: CXCR Inhibition The CXCR4 inhibitor AMD070 hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) and the CXCR7 inhibitor ACT-1004-1239 (MedChemExpress, Monmouth Junction, NJ, USA) were diluted to 2 mg/mL and 5 mg/mL with dimethyl sulfoxide (DMSO), respectively, according to manufacturer protocols.

Techniques: Inhibition, Control

Journal: OALib

Article Title: The Complementary Roles of CXCR4 and CXCR7 in Melanoma Migration

doi: 10.4236/oalib.1112617

Figure Lengend Snippet: Figure 3. CXCR4 and 7 inhibition decreases melanoma cell chemokinesis. Box plot graphs show how melanoma migration decreased after CXCR4 and CXCR7 inhibition. Data from 4 experiments was normalized to the control values. The number of asterisks present identifies ANOVA significant p-values and are in comparison to the Control + DMSO values. There was no significant difference between the CXCR4 and CXCR7 inhibitors. ***p < 0.001, ****p < 0.0001. N = 4. These results with the two inhibitors match what we know about SDF1 binding to CXCR4/7. Experimental studies on the thermodynamics and kinetics of recep- tor interactions with SDF1 have shown that CXCR7 has a higher affinity for SDF1 than CXCR4 [32]. Our structural modeling of the CXCR4:SDF1 and CXCR7:SDF1 complexes and their relaxation in the lipid bilayer environment; the binding free energy analysis of these interaction partners is shown in Appendix 2, along with experimental Kd values. We find that CXCR7 binds with a stronger affinity to SDF1 (more negative DG) than CXCR4, which is consistent with the above exper- imental observations. The structures provide a biophysical basis for CXCR7,

Article Snippet: CXCR Inhibition The CXCR4 inhibitor AMD070 hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) and the CXCR7 inhibitor ACT-1004-1239 (MedChemExpress, Monmouth Junction, NJ, USA) were diluted to 2 mg/mL and 5 mg/mL with dimethyl sulfoxide (DMSO), respectively, according to manufacturer protocols.

Techniques: Inhibition, Migration, Control, Comparison, Binding Assay

Journal: OALib

Article Title: The Complementary Roles of CXCR4 and CXCR7 in Melanoma Migration

doi: 10.4236/oalib.1112617

Figure Lengend Snippet: Figure 4. Effect of CXCR4/7 KD in melanoma wound healing. Bar graphs showing progressive healing of melanoma wounds at each time point over an 18-hour time frame for each set of transfections. The data starts at 0% of the wound area healed at 0 hr, and as time progresses, cells invade the wound and make the area smaller, healing the wound. Normal B16-F10 cells were compared to A. Scramble siRNA, B. CXCR4 siRNA, C. CXCR7 siRNA and D. CXCR4 and CXCR7 siRNAs. The only significant difference observed was between our control cells and CXCR4 and CXCR7 siRNA knockdown cells in the presence of SDF1. *p < 0.05.

Article Snippet: CXCR Inhibition The CXCR4 inhibitor AMD070 hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) and the CXCR7 inhibitor ACT-1004-1239 (MedChemExpress, Monmouth Junction, NJ, USA) were diluted to 2 mg/mL and 5 mg/mL with dimethyl sulfoxide (DMSO), respectively, according to manufacturer protocols.

Techniques: Transfection, Control, Knockdown

Journal: OALib

Article Title: The Complementary Roles of CXCR4 and CXCR7 in Melanoma Migration

doi: 10.4236/oalib.1112617

Figure Lengend Snippet: Figure 5. CXCR4/7 KD decreases melanoma cell migration. Box plot graphs show how melanoma cells decrease migration rates after CXCR4 and CXCR7 KD. Data from 5 experiments was normal- ized to the control values. ANOVA significant p-values are identified by the number of asterisks present: *p < 0.00001. Cell velocity is one of the critical indicators of increased/decreased cell migra- tion and is necessary for successful metastasis. We wanted to determine if CXCR4/7 influences melanoma migration. B16-F10 cells were transfected with scramble, CXCR4 KD, CXCR KD, or a combination of CXCR4 and 7 KD siRNA to measure a change. B16-F10 cells were then plated either in the presence of or without SDF1. Cell velocity was measured by manually tracking at least 15 cells at a time from each experiment using an ImageJ plug-in. The formula used to meas- ure velocity in ImageJ was the following:

Article Snippet: CXCR Inhibition The CXCR4 inhibitor AMD070 hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) and the CXCR7 inhibitor ACT-1004-1239 (MedChemExpress, Monmouth Junction, NJ, USA) were diluted to 2 mg/mL and 5 mg/mL with dimethyl sulfoxide (DMSO), respectively, according to manufacturer protocols.

Techniques: Migration, Control, Transfection

Journal: OALib

Article Title: The Complementary Roles of CXCR4 and CXCR7 in Melanoma Migration

doi: 10.4236/oalib.1112617

Figure Lengend Snippet: Figure 6. CXCR4/7 KD decreases B16-F10 melanoma cell velocity. Graphs represent the velocity, in µm per second, after melanoma transfection with A. CXCR4 siRNA, B. CXCR7 siRNA, and C. CXCR4 and CXCR7 siRNAs. siRNAs were compared to untreated and scrambled random siRNA. Significant p-values are identified by the number of asterisks present: *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001.

Article Snippet: CXCR Inhibition The CXCR4 inhibitor AMD070 hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) and the CXCR7 inhibitor ACT-1004-1239 (MedChemExpress, Monmouth Junction, NJ, USA) were diluted to 2 mg/mL and 5 mg/mL with dimethyl sulfoxide (DMSO), respectively, according to manufacturer protocols.

Techniques: Transfection

Journal: OALib

Article Title: The Complementary Roles of CXCR4 and CXCR7 in Melanoma Migration

doi: 10.4236/oalib.1112617

Figure Lengend Snippet: Figure 7. CXCR4/7 KD + SDF1 decreases B16-F10 melanoma cell velocity. Graphs repre- sent the velocity, in µm per second, after melanoma transfection with A. CXCR4 siRNA, B. CXCR7 siRNA, and C. CXCR4 and CXCR7 siRNAs and the addition of SDF1. siRNAs were compared to untreated + SDF1 and scramble random siRNA +SDF1. Significant p-values are identified by the number of asterisks present: *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001.

Article Snippet: CXCR Inhibition The CXCR4 inhibitor AMD070 hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) and the CXCR7 inhibitor ACT-1004-1239 (MedChemExpress, Monmouth Junction, NJ, USA) were diluted to 2 mg/mL and 5 mg/mL with dimethyl sulfoxide (DMSO), respectively, according to manufacturer protocols.

Techniques: Transfection

Journal: OALib

Article Title: The Complementary Roles of CXCR4 and CXCR7 in Melanoma Migration

doi: 10.4236/oalib.1112617

Figure Lengend Snippet: Figure 8. CXCR7 KD decreases melanoma cell size. A) Images of rat B16-F10 melanoma cell line stained with phalloidin-488 after siRNA transfection for CXCR4 or CXCR7. Arrows in CXCR7 KD point to long stress fibers in melanoma cells. B) We plotted the cells’ cell size area after transfection. The ** shows the significant reduction of CXCR after siRNA (Anova less than p < 0.0001). The number below boxplots corresponds to many cells traced (N = 100).

Article Snippet: CXCR Inhibition The CXCR4 inhibitor AMD070 hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) and the CXCR7 inhibitor ACT-1004-1239 (MedChemExpress, Monmouth Junction, NJ, USA) were diluted to 2 mg/mL and 5 mg/mL with dimethyl sulfoxide (DMSO), respectively, according to manufacturer protocols.

Techniques: Staining, Transfection

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: AMD3465 (hexahydrobromide) rescues the MG63 osteoblasts against the apoptosis induced by high glucose.

doi: 10.1016/j.biopha.2021.111476

Figure Lengend Snippet: Fig. 1. The effect of AMD3465 on the proliferation of MG63 cells mediated by high glucose. MG-63 cells were divided into three groups, including normal- glucose group (NC, 5.5 mmol/L glucose), high-glucose group (HG, 20 mmol/L glucose) and AMD3465 treaded group (HG+AMD3465, 20 mmol/L glucose plus 10 μmol/L AMD3465). The proliferation of MG63 cells was confirmed by CCK8 assay (A) at 1, 2 and 3 days after culture. The clonality of MG63 cells was measured by clone formation assay (B) at two weeks after culture. The results of the clone formation assay were captured with a camera. No magnification. (C) Western blot was used to detect the expression of cell cycle-related proteins in MG63 cells. (D) The quantitation of western blot. All experiments were independently carried out in 3 replicates. Note: Compared with NC, *P < 0.05; Compared with HG, #P < 0.05.

Article Snippet: MG-63 cells were divided into three groups, including normalglucose group (NC, α-MEM medium, 5.5 mmol/L glucose), highglucose group (HG, addition of glucose (14.5 mM) to the α-MEM medium, a final concentration of 20 mmol/L glucose) and AMD3465 (Med Chem Express) treaded group (HG+AMD3465, 20 mmol/L glucose plus 10 μmol/L AMD3465).

Techniques: CCK-8 Assay, Tube Formation Assay, Western Blot, Expressing, Quantitation Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: AMD3465 (hexahydrobromide) rescues the MG63 osteoblasts against the apoptosis induced by high glucose.

doi: 10.1016/j.biopha.2021.111476

Figure Lengend Snippet: Fig. 2. The effect of AMD3465 on the migration and invasion of MG63 cells mediated by high glucose. MG-63 cells were divided into three groups, including normal-glucose group (NC, 5.5 mmol/L glucose), high-glucose group (HG, 20 mmol/L glucose) and AMD3465 treaded group (HG+AMD3465, 20 mmol/L glucose plus 10 μmol/L AMD3465). (A) The ability of cell migration was detected by scratch test at 0 and 24 h after culture. (B) Transwell assay was performed to confirm the cell invasion at 24 h after culture. All experiments were independently carried out in 3 replicates. Scale bar, 100 µm. Note: Compared with NC, *P < 0.05. Compared with HG, #P < 0.05.

Article Snippet: MG-63 cells were divided into three groups, including normalglucose group (NC, α-MEM medium, 5.5 mmol/L glucose), highglucose group (HG, addition of glucose (14.5 mM) to the α-MEM medium, a final concentration of 20 mmol/L glucose) and AMD3465 (Med Chem Express) treaded group (HG+AMD3465, 20 mmol/L glucose plus 10 μmol/L AMD3465).

Techniques: Migration, Transwell Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: AMD3465 (hexahydrobromide) rescues the MG63 osteoblasts against the apoptosis induced by high glucose.

doi: 10.1016/j.biopha.2021.111476

Figure Lengend Snippet: Fig. 3. AMD3465 inhibits the high glucose-mediated apoptosis. MG-63 cells were divided into three groups, including normal-glucose group (NC, 5.5 mmol/L glucose), high-glucose group (HG, 20 mmol/L glucose) and AMD3465 treaded group (HG+AMD3465, 20 mmol/L glucose plus 10 μmol/L AMD3465). (A) The apoptosis of MG63 cells was analyzed using cell flow-cytometry at 24 h after culture. (B) Cell apoptosis was observed by Hoechst and PI double staining. (C) The expression of apoptosis-related proteins was detected by Western blotting. All experiments were independently carried out in 3 replicates. Note: Compared with NC, *P < 0.05; Compared with HG, #P < 0.05.

Article Snippet: MG-63 cells were divided into three groups, including normalglucose group (NC, α-MEM medium, 5.5 mmol/L glucose), highglucose group (HG, addition of glucose (14.5 mM) to the α-MEM medium, a final concentration of 20 mmol/L glucose) and AMD3465 (Med Chem Express) treaded group (HG+AMD3465, 20 mmol/L glucose plus 10 μmol/L AMD3465).

Techniques: Flow Cytometry, Double Staining, Expressing, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: AMD3465 (hexahydrobromide) rescues the MG63 osteoblasts against the apoptosis induced by high glucose.

doi: 10.1016/j.biopha.2021.111476

Figure Lengend Snippet: Fig. 4. AMD3465 inhibits the high glucose-induced expression of MMPs. MG-63 cells were divided into three groups, including normal-glucose group (NC, 5.5 mmol/L glucose), high-glucose group (HG, 20 mmol/L glucose) and AMD3465 treaded group (HG+AMD3465, 20 mmol/L glucose plus 10 μmol/L AMD3465). (A) The expression of SDF-1, CXCR4 and MMPS proteins was detected by Western blotting. (B) Quantitative results of western blot. All experiments were independently carried out in 3 replicates. Note: Compared with NC, *P < 0.05; Compared with HG, #P < 0.05.

Article Snippet: MG-63 cells were divided into three groups, including normalglucose group (NC, α-MEM medium, 5.5 mmol/L glucose), highglucose group (HG, addition of glucose (14.5 mM) to the α-MEM medium, a final concentration of 20 mmol/L glucose) and AMD3465 (Med Chem Express) treaded group (HG+AMD3465, 20 mmol/L glucose plus 10 μmol/L AMD3465).

Techniques: Expressing, Western Blot