Journal: International journal of oncology

Article Title: uPAR and cathepsin B knockdown inhibits radiation-induced PKC integrated integrin signaling to the cytoskeleton of glioma-initiating cells.

doi: 10.3892/ijo.2012.1496

Figure Lengend Snippet: Figure 3. Effect of pUC and radiation alone and in combination on PKC and integrin levels in U251 and 5310 non-GICs and GICs. A, Western blot analysis of the cell lysates of U251 and 5310 non-GICs and GICs showing the protein expression levels of PKC θ, PKC δ, pPKC θ/δ, PKC ζ, integrin β1, integrin α2, and integrin α5 after treatment with pUC and radiation alone or in combination. B, Total protein (300 µg) was collected from pSV, pUC, pSV + 10 Gy and pUC + 10 Gy samples of U251 and 5310 non-GICs and GICs and immunoprecipitated with integrin β1 antibody (2 µg) and protein A plus G agarose beads (20 µg) overnight at 4˚C. The precipitates were washed with lysis buffer and the integrin β1 pulled down protein was immunoblotted for pPKC θ/δ and PKC ζ. C, Co-localization of PKC ζ and integrin β1 was carried out with pSV and pUC with and without 10 Gy (24 h for non-GICs and 48 h for GICs). The cells were allowed to migrate on 4-well chamber slides for about 16 h after growing them in Ibidi culture inserts for 24 h after treatments. The cells were fixed, stained with primary antibody overnight at 4˚C, counter-stained with species-specific Alexa Fluor-conjugated secondary antibodies, nuclear stained with DAPI, mounted, and imaged under a confocal microscope. Arrows indicating the co-localization of PKC ζ and integrin β1.

Article Snippet: U251 and 5310 non-GICs or GICs were transfected with scrambled vector (pSV) or a bicistronic construct of uPAR and cathepsin B (pCU) or siRNA against integrin β1 (SCBT, Santa Cruz, CA).

Techniques: Western Blot, Expressing, Immunoprecipitation, Lysis, Staining, Microscopy

Journal: International journal of oncology

Article Title: uPAR and cathepsin B knockdown inhibits radiation-induced PKC integrated integrin signaling to the cytoskeleton of glioma-initiating cells.

doi: 10.3892/ijo.2012.1496

Figure Lengend Snippet: Figure 4. PKCs regulate the ECM-integrin interaction signal and vice versa in U251 and 5310 non-GICs and GICs. A, SDS-PAGE was conducted with the cell lysates of control, DMSO-treated, rottlerin (200 µM) and integrin β1 siRNA-treated U251 and 5310 non-GICs and GICs grown in non-coated and collagen-coated culture dishes. The cell lysates were immunoblotted with PKC θ, PKC δ, pPKC θ/δ, PKC ζ, integrin β1 and integrin α2. B, SDS-PAGE was conducted with the cell lysates of control, DMSO-treated, rottlerin (200 µM) and integrin β1 siRNA-treated U251 and 5310 non-GICs and GICs grown in non-coated and fibronectin- coated culture dishes. The cell lysates were immunoblotted for PKC θ, PKC δ, pPKC θ/δ, PKC ζ, integrin β1 and integrin α5.

Article Snippet: U251 and 5310 non-GICs or GICs were transfected with scrambled vector (pSV) or a bicistronic construct of uPAR and cathepsin B (pCU) or siRNA against integrin β1 (SCBT, Santa Cruz, CA).

Techniques: SDS Page, Control

Journal: International journal of oncology

Article Title: uPAR and cathepsin B knockdown inhibits radiation-induced PKC integrated integrin signaling to the cytoskeleton of glioma-initiating cells.

doi: 10.3892/ijo.2012.1496

Figure Lengend Snippet: Figure 5. Depletion of uPAR and cathepsin B inhibits PKC/integrin signaling to FAK and the cytoskeletal molecules. A, Cell lysates from U251 and 5310 non-GICs and GICs were extracted using RIPA buffer after treatment with pUC and radiation alone or in combination, and western blot analysis was performed to determine the expression levels of FAK, pFAK (Tyr397), pFAK (Tyr925), vinculin, and α-actinin using their specific antibodies. GAPDH was used as a loading control. B, The total cell lysates of U251 and 5310 non-GICs and GICs as described above were immunoprecipitated with FAK antibody (2 µg). The protein precipitates were washed with lysis buffer and incubated with 1X loading dye at 90˚C for 10 min. SDS-PAGE was conducted and western blotting was performed with vinculin and α-actinin antibodies as described in Materials and methods.

Article Snippet: U251 and 5310 non-GICs or GICs were transfected with scrambled vector (pSV) or a bicistronic construct of uPAR and cathepsin B (pCU) or siRNA against integrin β1 (SCBT, Santa Cruz, CA).

Techniques: Western Blot, Expressing, Control, Immunoprecipitation, Lysis, Incubation, SDS Page

Journal: International journal of oncology

Article Title: uPAR and cathepsin B knockdown inhibits radiation-induced PKC integrated integrin signaling to the cytoskeleton of glioma-initiating cells.

doi: 10.3892/ijo.2012.1496

Figure Lengend Snippet: Figure 7. Effect of pUC and radiation treatment on pre-established intracranial tumors. U251 non-GICs and GICs were implanted intracranially into nude mice and treated with 450 µg of pUC seven days after implantation. Radiation treatment was given in two doses (5 Gy at days 8 and 10). When chronic symptoms were observed, the mice were sacrificed, and their brains were collected and embedded in paraffin. Paraffin-embedded sections were deparaffinized, antigen retrieved, and co-localization studies were carried out. The brain sections were incubated overnight with primary antibodies in 10% goat serum at 4˚C in a humidified chamber, counterstained with Alexa Fluor-conjugated secondary antibodies, and incubated with DAPI for nuclear staining before mounting. A, Co-localization of integrin β1 (green), pPKC θ/δ (red) and DAPI (blue) in the paraffin sections of the nude mice established with U251 non-GICs and GICs and treated with shRNA and radiation alone or in combination. B, Interaction between integrin β1 (green) and PKC ζ (red) in the various in vivo combination treatments. DAPI (blue) was used for nuclear staining. C, In vivo brain sections of immunocompromised mice implanted with U251 non-GICs and GICs were labeled with FAK (green) and α-actinin (red) and processed for immunofluorescence. The sections were incubated with DAPI for a brief period of time for nuclear staining. D, Mock, pUC- treated, irradiated, and pUC + irradiated brain sections of the nude mice pre-established with U251 non-GICs and GICs were immunoprocessed and labeled with FAK (green), vinculin (red) and DAPI (blue) in order to observe the co-localization of FAK and vinculin in in vivo samples.

Article Snippet: U251 and 5310 non-GICs or GICs were transfected with scrambled vector (pSV) or a bicistronic construct of uPAR and cathepsin B (pCU) or siRNA against integrin β1 (SCBT, Santa Cruz, CA).

Techniques: Incubation, Staining, shRNA, In Vivo, Labeling, Immunofluorescence, Irradiation

Journal: International Journal of Molecular Sciences

Article Title: Integrin β1 Promotes Pancreatic Tumor Growth by Upregulating Kindlin-2 and TGF-β Receptor-2

doi: 10.3390/ijms221910599

Figure Lengend Snippet: Integrin α5 and β1 are associated with pancreatic cancer patient survival. Integrin β1 and α5 mRNA levels in primary tumors were compared with normal tissue using TCGA data. ( A ) Primary tumors have higher gene expression of integrin β1 than normal tissues (# p < 0.0001, n = 165 for normal and n = 178 for tumors) red lines indicate the mean and SD of the values. ( B ) Increased expression of integrin β1 (red lines) associates with lower survival in patients as depicted by KM plot p < 0.001. ( C ) Integrin α5 also has higher gene expression than normal tissues (# p < 0.0001, n = 165 for normal and n = 183 for tumors). ( D ) Integrin α5 did not show a clear relationship with patient survival p > 0.05.

Article Snippet: Integrin β1-KO MIA, PaCa-2 cell lines were established using Integrin β1 CRISPR/Cas9 knock out Plasmids (sc-421171-NIC-2: Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Gene Expression, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Integrin β1 Promotes Pancreatic Tumor Growth by Upregulating Kindlin-2 and TGF-β Receptor-2

doi: 10.3390/ijms221910599

Figure Lengend Snippet: The deletion of the β1 integrin gene from MIA PaCa-2 cells decreases tumor growth on the 3D matrix. ( A ) Tumors formed by MIA PaCa-2 cells expressing the integrin β1 gene and cells where the integrin β1 gene removed were stained for F-Actin (phalloidin, green) to estimate the size of the tumor. Scale bar 20 µm. ( B ) 3D projection of z stack indicate the tumor size. ( C ) The projection of the intensity of actin staining (view 2.5D) and (D) Tumor volumes were measured after the acquisition of z-stack slices with confocal microscope. These images were processed using ImageJ software to determine the volume. One-way ANOVA was performed with Bonferroni’s test to compare between groups. * indicates a statistically significant difference between WT and Clone 2, n = 12, p < 0.001). # indicates the statistically significant difference between groups WT and Clone 4 n = 12, p < 0.001. Data shown are mean ± SEM, unit: µm 3 .

Article Snippet: Integrin β1-KO MIA, PaCa-2 cell lines were established using Integrin β1 CRISPR/Cas9 knock out Plasmids (sc-421171-NIC-2: Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Staining, Microscopy, Software

Journal: International Journal of Molecular Sciences

Article Title: Integrin β1 Promotes Pancreatic Tumor Growth by Upregulating Kindlin-2 and TGF-β Receptor-2

doi: 10.3390/ijms221910599

Figure Lengend Snippet: Integrin β1 deletion decreased the cellular adhesion of MIA PaCa-2 cells to vitronectin and fibronectin. ( A ) Integrin β1 expressing MIA PaCa-2 and integrin β1 KO clones were immunoblotted by anti-integrin β1 antibody. GAPDH was used as the loading control. ( B ) Cell lysate was immunoblotted with integrin α5 antibodies. GAPDH was used as the loading control. (C) Quantification of integrin α5 expression normalized to GAPDH, mean ± SEM from three independent experiments ( n = 3). One-way ANOVA performed with Bonferroni’s test indicated a statistically significant decrease in cell adhesion, * p < 0.001 between WT and clone2, # p < 0.001 between WT and clone4. ( D ) The expression of integrin α5 mRNA by RT-PCR. ΔΔCt values normalized with to 18s n = 3, * p < 0.001 between WT and clone2, # p < 0.001 between WT and clone4. ( E ) Percentage change in cellular adherence of integrin β1 KO to fibronectin (5 µg/mL) compared to the integrin β1 expressing cells, n = 3, * p < 0.01 between WT and clone2, # p < 0.01 between WT and clone4. ( F ) Comparison of cellular adhesion between integrin β1 expressing MIA PaCa-2 cells and integrin β1 KO cells to vitronectin (5 µg/mL) n = 3, * p < 0.01 between WT and clone2, # p < 0.01 between WT and clone4.

Article Snippet: Integrin β1-KO MIA, PaCa-2 cell lines were established using Integrin β1 CRISPR/Cas9 knock out Plasmids (sc-421171-NIC-2: Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Clone Assay, Control, Reverse Transcription Polymerase Chain Reaction, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Integrin β1 Promotes Pancreatic Tumor Growth by Upregulating Kindlin-2 and TGF-β Receptor-2

doi: 10.3390/ijms221910599

Figure Lengend Snippet: MIA PaCa-2 integrin β1 KO cells have decreased spreading ability compared to integrin β1 expressing cells. Cells were plated on either fibronectin or vitronectin coated plates for 4 h and the cells were stained with actin. ( A ) Actin staining of cell spread on 5 µg/mL fibronectin visualized by immunostaining (F-actin, phalloidin, green; and nucleus, DAPI, blue). Scale bar 20 µm. ( B ) Actin staining of cell spread on vitronectin visualized by immunostaining (Green: F-actin using phalloidin, and Blue: nucleus DAPI). Scale bar 20 µm. ( C , D ) Quantitative assessment of cell spreading in fibronectin (area: µm 2 ). One-way ANOVA was performed with Bonferroni’s post-hoc test to compare between groups. * Indicates a statistically significant difference between groups (integrin β1 expressing MIA PaCa-2 and integrin β1 KO Clone 2) ( n = 40, p < 0.001). # Shows a statistically significant difference between groups (integrin β1 expressing MIA PaCa-2 and integrin β1 KO Clone 4) ( n = 40, p < 0.001). Data shown are mean ± SEM, unit: µm 2 .

Article Snippet: Integrin β1-KO MIA, PaCa-2 cell lines were established using Integrin β1 CRISPR/Cas9 knock out Plasmids (sc-421171-NIC-2: Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Staining, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Integrin β1 Promotes Pancreatic Tumor Growth by Upregulating Kindlin-2 and TGF-β Receptor-2

doi: 10.3390/ijms221910599

Figure Lengend Snippet: Impaired focal adhesion contributes to less cell spreading in MIA PaCa-2 integrin β1 KO cells ( A ) Phospho-paxillin immunostaining shows the decrease in the phosphorylation status of paxillin in MIA PaCa-2 and integrin β1 KO cells. Visualized by immunostaining F-actin, phalloidin, green, p-paxillin, Alexa flour 555, red). Scale bar 20 µm. ( B ) Immunoblotting confirmed the downregulation of p-paxillin. Normalized p-paxillin expression with GAPDH shows a significant decrease in expression of p-paxillin. ( C ) No significant change in paxillin expression normalized with GAPDH between WT and clones. One-way ANOVA was performed by Bonferroni’s test to compare between groups ( n = 3). * indicates a statistically significant difference between groups (integrin β1 expressing MIA PaCa-2 and integrin β1 KO cells Clone 2, p < 0.001). # Shows a statistically significant difference between groups (integrin β1 expressing MIA PaCa-2 and integrin β1 KO cells Clone 4, p < 0.001).

Article Snippet: Integrin β1-KO MIA, PaCa-2 cell lines were established using Integrin β1 CRISPR/Cas9 knock out Plasmids (sc-421171-NIC-2: Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Immunostaining, Phospho-proteomics, Western Blot, Expressing, Clone Assay

Journal: International Journal of Molecular Sciences

Article Title: Integrin β1 Promotes Pancreatic Tumor Growth by Upregulating Kindlin-2 and TGF-β Receptor-2

doi: 10.3390/ijms221910599

Figure Lengend Snippet: Integrin β1 deletion downregulated Kindlin-2 and TGFβ receptor 2 expression. ( A ) Cell lysate (30 µg/lane) from integrin β1 expressing MIA PaCa-2 and integrin β1 KO clones are immunoblotted for kindlin-2. Normalized protein expression with GAPDH shows a decrease in kindlin-2 expression in clones compared to the normal MIA PaCa-2 cells. ( B ) The mRNA expression quantified by RT-PCR analysis shows a significant decrease in the expression of kindlin-2 by the deletion of integrin β1. ( C ) Lysates were immune blotted with antibody recognizing TGFβ receptor 2. GAPDH is used as the loading control. Normalized protein expression shows downregulation of TGFβ receptor 2. ( D ) The mRNA expression quantified by RT-PCR analysis shows a significant decrease in the expression of TGFβ receptor 2 by the deletion of integrin β1. One-way ANOVA was performed by Bonferroni’s test to compare integrin β1 expressing MIA PaCa-2 and integrin β1 KO clones indicate a statistically significant difference n = 3, * WT and Clone 2 p < 0.001; # WT and Clone 4 p < 0.001. Quantification is expressed as mean ± SEM.

Article Snippet: Integrin β1-KO MIA, PaCa-2 cell lines were established using Integrin β1 CRISPR/Cas9 knock out Plasmids (sc-421171-NIC-2: Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Clone Assay, Reverse Transcription Polymerase Chain Reaction, Control

Journal: International Journal of Molecular Sciences

Article Title: Integrin β1 Promotes Pancreatic Tumor Growth by Upregulating Kindlin-2 and TGF-β Receptor-2

doi: 10.3390/ijms221910599

Figure Lengend Snippet: Integrin β1 deletion downregulated Smad2/3 phosphorylation. ( A ) Cell lysate from integrin β1 expressing MIA PaCa-2 and integrin β1 KO clones is immunoblotted for p-Smad2/3. ( A ) Immunoblotting of p-Smad2/3 shows inhibited downstream cell signaling pathways. Normalized P Smad-2/3 is shown using GAPDH as control. ( B ) Total expression of Smad-2/3 shows no significant change in the expression of Smad-2/3. One-way ANOVA performed with Bonferroni’s test to compare integrin β1 expressing MIA PaCa-2 and integrin β1 KO clones indicates a statistically significant difference n = 3, * WT and Clone 2 p < 0.001; # WT and Clone 4 p < 0.001. Quantification is expressed as mean ± SEM.

Article Snippet: Integrin β1-KO MIA, PaCa-2 cell lines were established using Integrin β1 CRISPR/Cas9 knock out Plasmids (sc-421171-NIC-2: Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Phospho-proteomics, Expressing, Clone Assay, Western Blot, Protein-Protein interactions, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: The Sall2 transcription factor promotes cell migration regulating focal adhesion turnover and integrin β1 expression

doi: 10.3389/fcell.2022.1031262

Figure Lengend Snippet: Sall2 promotes integrin β1 expression. (A–C) Representative blots of integrin β1 expression, and densitometric quantification, from Sall2 +/+ and Sall2 −/− iMEFs after 15 min of nocodazole wash-out treatment (NZ w-o) (A) , 10 min of cell spreading (SPD) on FN (B) , and in normal growth conditions (NGC) (C) . (D) Representative blot of integrin β1 protein level, and densitometry, from the Tet-On Sall2 iMEFs inducible model after 15 min of NZ w-o. (E) Flow cytometry analysis of the cell surface expression of integrin β1 from Sall2 +/+ and Sall2 −/− iMEFs. (F) Left, a representative blot of brain tissues from 6 to 8-week-old Sall2 +/+ and Sall2 −/− mice. Right, integrin β1 densitometry from brain tissues. The arrow indicates Sall2, and the asterisk corresponds to a nonspecific band. The arrowheads in (A,B) indicate cropped unrelated columns and subsequent splicing of the blot. α-tubulin was used as loading control in western blot analysis. (G) Western blot analysis of integrin β1 overexpression in Sall2 −/− iMEFs. (H) Left, representative phase-contrast images (10x) at 0 and 16 h of in vitro wounding from Sall2 −/− iMEFs overexpressing integrin β1. Right, quantification of wound closure (as a percentage) at 16 h from images obtained in Left. (I) same as in H, but for Sall2 +/+ iMEFs incubated with anti-β1 antibody. Data are expressed as mean ± SD from three independent experiments (* p = 0.01 to 0.05, ** p = 0.001 to 0.01;unpaired t -test).

Article Snippet: Puromycin dihydrochloride (sc-108071), normal mouse IgG2A (sc-3878), Integrin β1 (M-106, sc-8978), Integrin β1 (A-4, sc-374429), Integrin α4 (B-2, sc-376334), Integrin α6 (F-6, sc-374057), Calpain 2 (E-10, sc-373966), FAK (H-1, sc-1688), PTEN (A2B1, sc-7974), Talin (C-9, sc-365875), Integrin β3 (N-20, sc-6627) and Vinculin (7F9, sc-73614) antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, United States).

Techniques: Expressing, Flow Cytometry, Control, Western Blot, Over Expression, In Vitro, Incubation

Journal: Frontiers in Cell and Developmental Biology

Article Title: The Sall2 transcription factor promotes cell migration regulating focal adhesion turnover and integrin β1 expression

doi: 10.3389/fcell.2022.1031262

Figure Lengend Snippet: Sall2 transcriptionally regulates integrin β1. (A) Bigwig track from ITGB1 gene, according to reference from NCBI RefSeq genes and UCSC genes (ENCODE), from SALL2 wild-type (WT) and SALL2 knockout HEK293 cells (ΔSall2) ChIP-seq datasets. SALL2 binding enrichment and associated peaks in the promoter region, according with EPDnew Promoters ENCODE data are shown. H3K27Ac enhancer mark and DNase Clusters from ENCODE are also shown, plotted with the UCSC genome browser ( https://genome.ucsc.edu/ , hg19 track). (B) Schematic representation of mouse and human integrin β1 ( Itgb1/ITGB1 ) promoters. The putative SALL2/Sall2 binding sites are represented by black ovals. The transcription start site (+1) is represented by arrows. (C–E) Integrin β1 mRNA level from Sall2 +/+ and Sall2 −/− iMEFs were determined by real-time PCR after 15 min of nocodazole wash-out treatment (NZ w-o) (C) , 10 min of spreading (SPD) on FN (D) and under normal growth conditions (NGC) (E) . (F) Integrin β1 mRNA levels from the Tet-On Sall2 iMEFs inducible model after 15 min of NZ w-o. RNA polymerase II was used as a normalizer. (G) ITGB1 promoter activity in the absence (FLAG vector) and presence of SALL2 (FLAG_SALL2) was performed as described in the MATERIALS AND METHODS. Luciferase activity was measured from cell lysates and normalized to β-galactosidase activity, and promoter activity was expressed as relative luciferase units (R.L.U). pGL3 vector served as control. (H) Schematic representation of human ITGB1 promoter. Horizontal arrows indicate the location of primers used for qPCR in site-specific ChIP assays. (I) Chromatin from SALL2 KO HEK293 cells transfected with FLAG_SALL2 was immunoprecipitated 24 h after transfection using FLAG antibody. Specific genomic regions of the human ITGB1 promoter and a nonrelated promoter region (NRR) were analyzed by real-time PCR. Graphs show quantification of the amplified DNA for each immunoprecipitation relative to FLAG. Data are expressed as mean ± SD from three independent experiments (n.s, not significant, * p = 0.01 to 0.05, ** p = 0.001 to 0.01; unpaired t -test).

Article Snippet: Puromycin dihydrochloride (sc-108071), normal mouse IgG2A (sc-3878), Integrin β1 (M-106, sc-8978), Integrin β1 (A-4, sc-374429), Integrin α4 (B-2, sc-376334), Integrin α6 (F-6, sc-374057), Calpain 2 (E-10, sc-373966), FAK (H-1, sc-1688), PTEN (A2B1, sc-7974), Talin (C-9, sc-365875), Integrin β3 (N-20, sc-6627) and Vinculin (7F9, sc-73614) antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, United States).

Techniques: Knock-Out, ChIP-sequencing, Binding Assay, Real-time Polymerase Chain Reaction, Activity Assay, Plasmid Preparation, Luciferase, Control, Transfection, Immunoprecipitation, Amplification

Journal: Frontiers in Cell and Developmental Biology

Article Title: The Sall2 transcription factor promotes cell migration regulating focal adhesion turnover and integrin β1 expression

doi: 10.3389/fcell.2022.1031262

Figure Lengend Snippet: Proposed model of Sall2-dependent regulation of cell migration. In Sall2 +/+ iMEFs cells, (1) Sall2 promotes the integrin β1 mRNA expression by directly binding to and transactivating its promoter, (2) increasing the integrin β1 protein levels and its expression at the membrane surface, contributing in part to the integrin β1 cluster formation. (3) After integrin ligand-binding, actin-associated proteins like talin and vinculin bind to the integrin β1 and recruit focal adhesion kinase (FAK). (4) Once recruited in the adhesion site, FAK is autophosphorylated modulating FA assembly-disassembly dynamics. Conversely, in Sall2 −/− cells the same chain of events occurs but less favorably. In summary, Sall2 by promoting the FA dynamics favors cell detachment and subsequently cell migration.

Article Snippet: Puromycin dihydrochloride (sc-108071), normal mouse IgG2A (sc-3878), Integrin β1 (M-106, sc-8978), Integrin β1 (A-4, sc-374429), Integrin α4 (B-2, sc-376334), Integrin α6 (F-6, sc-374057), Calpain 2 (E-10, sc-373966), FAK (H-1, sc-1688), PTEN (A2B1, sc-7974), Talin (C-9, sc-365875), Integrin β3 (N-20, sc-6627) and Vinculin (7F9, sc-73614) antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, United States).

Techniques: Migration, Expressing, Binding Assay, Membrane, Ligand Binding Assay