Journal:

Article Title: Integrin ? v ? 3 requirement for VEGFR2-mediated activation of SAPK2/p38 and for Hsp90-dependent phosphorylation of focal adhesion kinase in endothelial cells activated by VEGF

doi:

Figure Lengend Snippet: VEGF-induced migration of HUVEC on vitronectin is mediated by VEGFR2 and integrin αvβ3. (A) HUVEC in suspension were either not incubated or incubated for 20 minutes with optimal concentrations or dilutions of anti-αvβ3 (1 μg/mL), anti-αvβ5 (1 μg/mL), or anti-β1 (1:1000) integrin antibodies. Cells were then plated in 96-well culture dishes precoated with 3 μg/mL vitronectin. After 60 minutes at 37°C, adhering cells were stained with crystal violet and quantified by optical density (550 nm). Data points represent means ± SD of triplicate samples from 3 different experiments. (B) HUVEC in suspension were either not preincubated or preincubated for 20 minutes with anti-αvβ3 (1 μg/mL), anti-αvβ5 (1 μg/mL), or anti-β1 (1:1000) integrin antibodies. (C) HUVEC in suspension were preincubated in the presence or absence of a VEGFR2 blocking (VEGFR2-ba 10 μg/mL, 20 minutes) or nonblocking antibody (VEGFR2-nba 10 μg/mL, 20 minutes). After treatments, in both B and C, cells were seeded on the upper part of a vitronectin-coated membrane in a modified Boyden chamber either containing or not containing VEGF (5 ng/mL) in the lower part. After 4 hours of migration, cells on the upper part of the membrane were scraped, and the cells on the lower part were stained with Mayer's hematoxylin. The cells of each well were counted in 5 fields at 100× magnification. Data points represent means from triplicate samples taken from at least 3 different experiments. VEGF, vascular endothelial growth factor; HUVEC, human umbilical vein endothelial cells; VEGFR2, VEGF receptor 2

Article Snippet: Function-blocking antibody against integrin β1 subunit (clone P4C10) was from Life Technology, Burlington, ON, Canada.

Techniques: Migration, Incubation, Staining, Blocking Assay, Modification

Journal:

Article Title: Integrin ? v ? 3 requirement for VEGFR2-mediated activation of SAPK2/p38 and for Hsp90-dependent phosphorylation of focal adhesion kinase in endothelial cells activated by VEGF

doi:

Figure Lengend Snippet: VEGF-induced SAPK2/p38 activation and FAK tyrosine phosphorylation require both VEGFR2 and integrin αvβ3. (A) Quiescent HUVEC on vitronectin or in suspension were either not pretreated or pretreated with a VEGFR2 blocking antibody (VEGFR2-ba 10 μg/mL, 20 minutes) and then were either treated or not treated with VEGF (5 ng/mL, 5 minutes). Cells were then extracted and subjected to SAPK2/p38 assay. Extracts were separated by SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was processed by Western blot for phospho p38 detection. The membrane was stripped and reprobed for total p38 to ensure equal protein loading. Data points represent means ± SD of duplicate samples from 2 separate experiments. Representative blots are shown. (B) Quiescent HUVEC were maintained on vitronectin or put in suspension for 20 minutes. Adhering cells were either not pretreated or pretreated with a VEGFR2 blocking antibody (VEGFR2-ba 10 μg/mL, 20 minutes) and then were treated with VEGF (5 ng/mL, 5 minutes). Cells in suspension were either not pretreated or pretreated with the VEGFR2 blocking antibody (VEGFR2-ba 10 μg/mL, 20 minutes), and VEGF (5 ng/mL) was added for the last 5 minutes of suspension. Cells were then extracted and subjected to FAK immunoprecipitation, separated on SDS-PAGE, and transferred to a nitrocellulose membrane. The membrane was processed for phosphotyrosine detection. The membrane was stripped and reprobed for total FAK to ensure equal protein loading. Data points represent means ± SD of duplicate samples from 2 separate experiments. Representative blots are shown. (C, D) Cells in suspension were either not pretreated or pretreated with anti-integrin αvβ3 antibody (1 μg/mL, 20 minutes) and then were plated on vitronectin for 60 minutes. After 55 minutes, cells were either not treated or treated with VEGF (5 ng/mL, 5 minutes). In C, cells were extracted and subjected to p38 assay as in A. In D, cells were extracted and subjected to FAK assay as in B. Data points represent means of triplicate samples. Representative blots are shown. VEGF, vascular endothelial growth factor; SAPK2/p38, stress-activated protein kinase-2/p38; FAK, focal adhesion kinase; HUVEC, human umbilical vein endothelial cells; VEGFR2, VEGF receptor 2; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis

Article Snippet: Function-blocking antibody against integrin β1 subunit (clone P4C10) was from Life Technology, Burlington, ON, Canada.

Techniques: Activation Assay, Blocking Assay, SDS Page, Western Blot, Immunoprecipitation, Polyacrylamide Gel Electrophoresis

Journal:

Article Title: Integrin ? v ? 3 requirement for VEGFR2-mediated activation of SAPK2/p38 and for Hsp90-dependent phosphorylation of focal adhesion kinase in endothelial cells activated by VEGF

doi:

Figure Lengend Snippet: VEGF-induced SAPK2/p38 activation and FAK tyrosine phosphorylation require clustering of integrin αvβ3. (A) Quiescent cells were put in suspension and were either not pretreated or pretreated for the last 10 minutes of suspension with 10 nM contortrostatin. Then, they were either not treated or treated with VEGF (5 ng/mL, last 5 minutes of suspension) in the presence or absence of the VEGFR2 blocking antibody (VEGFR2-ba 10 μg/mL). Cells were then extracted and processed for SAPK2/p38 assay as in Figure 3A. Data points represent means of triplicate samples for each condition. Results are representative of 2 different experiments. (B) Quiescent HUVEC maintained on vitronectin were left to adhere or were put in suspension for 20 minutes. Adhering cells were either not treated or treated with VEGF (5 ng/mL, 5 minutes). Cells in suspension were either not pretreated or pretreated with 10 nM contortrostatin for the last 10 minutes of suspension and were either not treated or treated with VEGF (5 ng/mL, last 5 minutes of suspension). Cells were then extracted and subjected to FAK phosphorylation assay as in Figure 3B. Data points represent means of duplicate samples. Results are representative of 2 different experiments. VEGF, vascular endothelial growth factor; SAPK2/p38, stress-activated protein kinase-2/p38; FAK, focal adhesion kinase; VEGFR2, VEGF receptor 2; HUVEC, human umbilical vein endothelial cells

Article Snippet: Function-blocking antibody against integrin β1 subunit (clone P4C10) was from Life Technology, Burlington, ON, Canada.

Techniques: Activation Assay, Blocking Assay, Phosphorylation Assay

Journal:

Article Title: Integrin ? v ? 3 requirement for VEGFR2-mediated activation of SAPK2/p38 and for Hsp90-dependent phosphorylation of focal adhesion kinase in endothelial cells activated by VEGF

doi:

Figure Lengend Snippet: Integrin αvβ3 mediates tyrosine phosphorylation of VEGFR2 in response to VEGF. (A, B) Quiescent HUVEC were maintained on vitronectin or put in suspension for 20 minutes. Adhering cells (A) were either not pretreated or pretreated with geldanamycin (GA, 1 μg/mL, 60 minutes) and then were either not treated or treated with VEGF (5 ng/mL, 5 minutes). For cells in suspension (B), contortrostatin (CN, 10 nM) was either not added or added for the last 10 minutes of suspension, and VEGF (5 ng/mL) was either not added or added for the last 5 minutes. Cells were then extracted and subjected to VEGFR2 immunoprecipitation. Thereafter, they were separated on sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane to detect phosphotyrosinated VEGFR2. The membrane was stripped and reprobed for total VEGFR2 to ensure equal protein loading. In the blank track (A, B), no cellular extract was added. Results are representative of 2 different experiments. VEGF, vascular endothelial growth factor; VEGFR2, VEGF receptor 2; HUVEC, human umbilical vein endothelial cells

Article Snippet: Function-blocking antibody against integrin β1 subunit (clone P4C10) was from Life Technology, Burlington, ON, Canada.

Techniques: Immunoprecipitation, Polyacrylamide Gel Electrophoresis

Journal: The Journal of Neuroscience

Article Title: Perisynaptic Chondroitin Sulfate Proteoglycans Restrict Structural Plasticity in an Integrin-Dependent Manner

doi: 10.1523/JNEUROSCI.2406-12.2012

Figure Lengend Snippet: List of primary reagents/antibodies used for immunofluorescence

Article Snippet: For studying the role of integrins in spine dynamics, slices were pretreated with a functional β1-integrin blocker (MAB1987Z; Millipore) or ACSF (control) overnight.

Techniques: Incubation

Journal: The Journal of Neuroscience

Article Title: Perisynaptic Chondroitin Sulfate Proteoglycans Restrict Structural Plasticity in an Integrin-Dependent Manner

doi: 10.1523/JNEUROSCI.2406-12.2012

Figure Lengend Snippet: CSPG digestion-induced enhancement of spine dynamics is dependent on β1-integrins. A, Representative confocal images of living YFP-positive CA1 pyramidal apical dendrites in the different treatment conditions. The white arrows indicate SHPs. B, ChABC-mediated enhancement of spine motility was blocked by pretreatment with a β1-integrin blocking antibody. C, Fluctuations in length over time for representative spines. The average spine length was not affected by ChABC treatment. D, CSPG digestion resulted in a doubling of the percentage of motile spines. Spines were considered motile when the respective motility index was higher than the average motility index of the control group. E, ChABC induced the formation of SHPs. Blocking of glutamatergic receptors failed to inhibit formation of SHPs, while pretreatment with a β1-integrin blocking antibody prevented their appearance. LtT, Long-term treatment; ACSF, artificial CSF microinjection; β1Ab, β1-integrin blocking antibody microinjection; β1Ab+ChABC, β1-integrin blocking antibody microinjection followed by ChABC treatment. ***p < 0.001, Student's t test. Error bars indicate SEM.

Article Snippet: For studying the role of integrins in spine dynamics, slices were pretreated with a functional β1-integrin blocker (MAB1987Z; Millipore) or ACSF (control) overnight.

Techniques: Blocking Assay

Journal: The Journal of Neuroscience

Article Title: Perisynaptic Chondroitin Sulfate Proteoglycans Restrict Structural Plasticity in an Integrin-Dependent Manner

doi: 10.1523/JNEUROSCI.2406-12.2012

Figure Lengend Snippet: CSPG digestion promotes activation of β1-integrins and phosphorylation of FAK. A, A higher density of active β1-integrins and phosphorylated FAK-positive puncta was detected within the CA1 stratum radiatum after ChABC treatment compared with control slices (active β1-int: control, 2.35 ± 0.27 puncta, 5 slices; ChABC, 4.81 ± 0.7 puncta, 5 slices; phospho-FAK: control, 5.5 ± 0.76 puncta, 10 slices; ChABC, 9.9 ± 0.93 puncta, 10 slices). ChABC treatment did not change the level of expression of native FAK and β1-integrin. Active β1-integrin puncta were significantly larger in treated slices compared with control slices (control, 0.28 ± 0.01 μm2, 5 slices; ChABC, 0.33 ± 0.01 μm2, 5 slices). B, Active β1-integrin-positive puncta were often associated with phosphorylated FAK-positive puncta in CA1 stratum radiatum after ChABC treatment. DAPI staining was used to visualize the CA1 strata. The white arrows indicate active β1-integrin-positive puncta (green) colocalizing with phosphorylated FAK-positive puncta (red). C, In ChABC-treated slices, phosphorylated FAK-positive puncta (red) were detected in association with YFP-positive dendritic spines (green) and the synaptic marker PSD-95 (blue). On average, 60.1 ± 2.09% of phosphorylated FAK puncta colocalized with PSD-95 puncta after CSPGs digestion compared with 41.28 ± 2.26% in control slices. D, High-magnification images show phosphorylated FAK-positive puncta (red) and PSD-95-positive puncta (blue) in the stratum radiatum of the CA1 field in control and ChABC-treated slices. **p < 0.01, ***p < 0.001, Student's t test. Error bars indicate SEM.

Article Snippet: For studying the role of integrins in spine dynamics, slices were pretreated with a functional β1-integrin blocker (MAB1987Z; Millipore) or ACSF (control) overnight.

Techniques: Activation Assay, Expressing, Staining, Marker

Journal: The Journal of Biological Chemistry

Article Title: Matrix Metalloproteinase-9 Regulates Survival of Neurons in Newborn Hippocampus *

doi: 10.1074/jbc.M111.297671

Figure Lengend Snippet: Inhibition of MMPs and laminin signaling regulate neuronal survival in vitro. Upper panel, MAP2 immunostaining of control (cnt), FN439-treated, laminin antibody (α-LM)-treated cultures, and cultures treated with both FN-439 and α-LM. Lower panel, quantification of neuron numbers (n = 10; means ± S.E.) Asterisks, p < 0.05 (one-way ANOVA). Neurons were incubated from DIV5 to DIV7. 50 μm FN439, 50 μg/ml α-LM, and 50 μg/ml anti-integrin β1 antibody (α-β1) were used.

Article Snippet: For function blocking, hamster anti-integrin β1 (50 mg/ml; BD Biosciences) and rabbit anti-laminin (50 mg/ml; Sigma) were used.

Techniques: Inhibition, In Vitro, Immunostaining, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Matrix Metalloproteinase-9 Regulates Survival of Neurons in Newborn Hippocampus *

doi: 10.1074/jbc.M111.297671

Figure Lengend Snippet: In vivo injection of FN-439 inhibits MMP activity and suppresses apoptosis. A, in situ zymography. FN-439 was injected at P2, and its effect was analyzed at P4. Left panel, CA3 region of control and FN-439-injected animal. DAPI staining (left) and gelatin-FITC (right). Right panel, fluorescence intensities of gelatin-FITC. CA3 regions of the control and FN-439-injected animals were compared (n = 5; means ± S.E.). Asterisk, p < 0.01 (Student's t test). B, immunostaining of laminin. Left panel, CA3 regions of the control and FN-439-treated animals are shown. Resolution of fluorescent intensities is 0–254 levels, with the color code for intensity shown as a scale bar. Right panel, intensity of laminin immunostaining signals (n = 6; means ± S.E.). Asterisk, p < 0.0001 (Student's t test). C, c-cas3+ neurons in the control, FN-439-injected and anti-integrin β1 antibody (α-β1)-injected animals were analyzed for CA1, CA3 pyramidal cells, and CA1 SO interneurons (n = 10; means ± S.E.). Asterisks, p < 0.05 (one-way ANOVA). cnt, control.

Article Snippet: For function blocking, hamster anti-integrin β1 (50 mg/ml; BD Biosciences) and rabbit anti-laminin (50 mg/ml; Sigma) were used.

Techniques: In Vivo, Injection, Activity Assay, In Situ, Zymography, Staining, Fluorescence, Immunostaining

Journal: The Journal of Biological Chemistry

Article Title: Matrix Metalloproteinase-9 Regulates Survival of Neurons in Newborn Hippocampus *

doi: 10.1074/jbc.M111.297671

Figure Lengend Snippet: As neurons mature, levels of MMP9 and laminin change, regulating critical survival signaling triggered by neuronal activity. In early postnatal neurons, levels of MMP9 that cleave laminin are high. As neurons mature, the levels of MMP9 decline, and laminin expression increases. Laminin-integrin signaling is required for depolarization-induced (L-type calcium-dependent) Ser(P)-473 Akt, which is critical for neuronal survival during the developmental period of neuronal death.

Article Snippet: For function blocking, hamster anti-integrin β1 (50 mg/ml; BD Biosciences) and rabbit anti-laminin (50 mg/ml; Sigma) were used.

Techniques: Activity Assay, Expressing

Journal: The Journal of Neuroscience

Article Title: Deleted in Colorectal Carcinoma and Differentially Expressed Integrins Mediate the Directional Migration of Neural Precursors in the Rostral Migratory Stream

doi: 10.1523/JNEUROSCI.22-09-03568.2002

Figure Lengend Snippet: Six integrins are differentially expressed in the rostral migratory stream. The α1 (A) and β8 (B) integrin subunits are expressed (arrows) from the anterior horn of the subventricular zone to the center of the olfactory bulb from P0 mice.C, The β1-integrin subunit is expressed in the RMS (arrows), blood vessels (BV), and the choroid plexus (CPX) of the lateral ventricle (LV) of a P2 mouse. D, The αv-integrin subunit is found in the RMS (arrows) from a P30 mouse. E, The β6-integrin subunit is expressed in the RMS (arrows) of a P15 mouse. F, The β3-integrin subunit is observed in P30 rat RMS (arrows). Scale bar, 1 mm. AOB, Accessory olfactory bulb; CC, corpus callosum; CX, cerebral cortex.

Article Snippet: The function-blocking anti-β1-integrin rat monoclonal antibody (clone 9EG7) was obtained from PharMingen (San Diego, CA); its specificity has been reported previously ( Lenter et al., 1993 ; Lenter and Vestweber, 1994 ).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Deleted in Colorectal Carcinoma and Differentially Expressed Integrins Mediate the Directional Migration of Neural Precursors in the Rostral Migratory Stream

doi: 10.1523/JNEUROSCI.22-09-03568.2002

Figure Lengend Snippet: Summary of the stage-specific expression of integrin and laminin subunits in the RMS. Tenascin-C is expressed along the sides of the RMS but not in the RMS itself.

Article Snippet: The function-blocking anti-β1-integrin rat monoclonal antibody (clone 9EG7) was obtained from PharMingen (San Diego, CA); its specificity has been reported previously ( Lenter et al., 1993 ; Lenter and Vestweber, 1994 ).

Techniques: Expressing

Journal: The Journal of Neuroscience

Article Title: Deleted in Colorectal Carcinoma and Differentially Expressed Integrins Mediate the Directional Migration of Neural Precursors in the Rostral Migratory Stream

doi: 10.1523/JNEUROSCI.22-09-03568.2002

Figure Lengend Snippet: Integrins mediate migration of neural precursors in the RMS. Brain slices from a P12 mouse were labeled with DiI and cultured in CCM1 medium with 5% horse serum for 5 hr in the presence of either control or anti-integrin antibodies. Three hours after addition of DiI, the migration was confirmed by fluorescence time lapse, and then a control or function-blocking anti-αv-integrin antibody was added for (Figure legend continued.) 5 hr, after which migration was recorded over the next 3 hr. A, Time-lapse sequence of three cells (a–c) in a slice migrating from the SVZ (bottom) toward the olfactory bulb (top) in the presence of a control antibody. Thearrow pointing to each cell shows the leading process, and the line shows the cell body. The interval between each image is 5 min. See Movie 1 (available at www.jneurosci.org).B, Graphical representation of the migration of the three cells (a–c) in A. Eachpoint represents the position of the cell body at 5 min time points. Note the unidirectional pathway and the bursts of rapid migration followed by slower meandering. C, Time-lapse sequence of images of seven cells (d–j) in a slice migrating from the SVZ (left) to the olfactory bulb (right) in the presence of an anti-αv-integrin antibody. Seven cells (d–j) are marked for reference. See Movie 2 (available at www.jneurosci.org). D, Graphical representation of the migration of the seven cells as described in C. The olfactory bulb is at theright. Note the inhibited migration. The interval between each image is 5 min. Scale bars, 50 μm.

Article Snippet: The function-blocking anti-β1-integrin rat monoclonal antibody (clone 9EG7) was obtained from PharMingen (San Diego, CA); its specificity has been reported previously ( Lenter et al., 1993 ; Lenter and Vestweber, 1994 ).

Techniques: Migration, Labeling, Cell Culture, Fluorescence, Blocking Assay, Sequencing

Journal: The Journal of Neuroscience

Article Title: Deleted in Colorectal Carcinoma and Differentially Expressed Integrins Mediate the Directional Migration of Neural Precursors in the Rostral Migratory Stream

doi: 10.1523/JNEUROSCI.22-09-03568.2002

Figure Lengend Snippet: Inhibited migration of RMS cells by function-blocking anti-integrin antibodies. Living brain slices were prepared from P3, P5, and P12 mice, and the cells were labeled with a small crystal of DiI placed on the center of the RMS. The slices were cultured in CCM1 medium supplemented with HEPES and 5% horse serum. The slices were preincubated with anti-integrin antibodies for 5 hr, and the migrating cells were traced by time-lapse recording for 3 hr. At P3, when α1- and β1-integrins are expressed, corresponding blocking antibodies reduced the migration speed. Anti-β3 antibody did not inhibit the migration significantly. At P5, when αv- and β1-integrins are expressed, antibodies against these integrins inhibited the migration speed. At P12, when αv-integrin is expressed, anti-αv-integrin antibody inhibited the speed as well; however, anti-α1 and -β3 antibodies did not inhibit the migration speed significantly, and they are not expressed at this stage. Each value represents the mean ± SD. Statistical analysis was performed by one-way ANOVA with Scheffé's multiple comparison procedure (significance of p < 0.01). The groups withasterisks do not differ from each other, nor do the nonmarked groups, but in all other comparisons, the differences are significant.

Article Snippet: The function-blocking anti-β1-integrin rat monoclonal antibody (clone 9EG7) was obtained from PharMingen (San Diego, CA); its specificity has been reported previously ( Lenter et al., 1993 ; Lenter and Vestweber, 1994 ).

Techniques: Migration, Blocking Assay, Labeling, Cell Culture

Journal: The Journal of Neuroscience

Article Title: Deleted in Colorectal Carcinoma and Differentially Expressed Integrins Mediate the Directional Migration of Neural Precursors in the Rostral Migratory Stream

doi: 10.1523/JNEUROSCI.22-09-03568.2002

Figure Lengend Snippet: Expression of DCC, neogenin, and netrin-1 and function of DCC in RMS migrations. A, Neogenin, a netrin-1 receptor, immunoreactivity coincides with the contour of RMS beginning at the anterior horn of the lateral ventricle (LV) and ending at the center of the olfactory bulb in a P0 mouse. B, The rostral part of the RMS strongly expresses the DCC protein, which is also present in the lateral olfactory tract (LOT) in P2 rats.AOB, Accessory olfactory bulb. C, Netrin-1 protein is expressed in the basal portion of olfactory mitral cells in the mitral cell layer (MCL) from an embryonic day 18 mouse. EPL, External plexiform layer;GCL, granule cell layer; GL, glomerular layer; ONL, olfactory nerve layer. (Figure legend continued.) D, Preabsorbed netrin-1 antibody (control) was prepared by coincubation of antibody and antigen peptide. The absorbed antibody did not show immunoreactivity. E, Sequence of 21 time-lapse images of a living slice from a P3 mouse treated with anti-DCC antibody as described in the legend to Figure ​Figure5.5. The olfactory bulb is located at the top. Three migrating cells (a–c) are indicated. Arrows indicate retracting leading process, and crossed arrows indicate processes pointing toward the anterior region of the subventricular zone (bottom). Note that the migration is no longer unidirectional, and that the processes form and retract frequently, which also distinguishes these migrations from those of normal cells shown in Figure ​Figure5.5. The interval between each image is 10 min. See Movie 3 (available atwww.jneurosci.org). F, Graphical representation of three migrating cells (a–c) shown in E. Cell bodies are tracked as described in Figure ​Figure5,5, and the time interval is 5 min. S, Start point; E, end point for cell b. See Movie 3 (available at www.jneurosci.org).G, Inhibited migration of cells by anti-DCC function-blocking antibodies. Living brain slices were prepared from P3 mice, and then the cells were labeled with DiI placed at the center or end of the RMS. The slices were cultured in CCM1 medium with HEPES and 5% horse serum with or without anti-DCC antibody for 5 hr, and the migration was observed by time-lapse recording. Each value represents the mean ± 1 SD. Statistical analysis was performed by one-way ANOVA with Scheffé's multiple comparison procedure (significance of p < 0.01). The control groups without anti-DCC antibody differ from every other group with anti-DCC. The groups withasterisks do not differ from each other, nor do the nonmarked groups, but in all other comparisons, the differences are significant. In contrast to cell treated with anti-integrin antibodies (Fig. ​(Fig.6),6), these cells show a larger net translocation. Scale bars:A, B, 1 mm; C, D, 100 μm;E, 50 μm.

Article Snippet: The function-blocking anti-β1-integrin rat monoclonal antibody (clone 9EG7) was obtained from PharMingen (San Diego, CA); its specificity has been reported previously ( Lenter et al., 1993 ; Lenter and Vestweber, 1994 ).

Techniques: Expressing, Sequencing, Migration, Blocking Assay, Labeling, Cell Culture, Translocation Assay

Journal: The Journal of Neuroscience

Article Title: Deleted in Colorectal Carcinoma and Differentially Expressed Integrins Mediate the Directional Migration of Neural Precursors in the Rostral Migratory Stream

doi: 10.1523/JNEUROSCI.22-09-03568.2002

Figure Lengend Snippet: Diagram depicting a working hypothesis for the roles of DCC and netrin and integrin in the migration of neural precursors from the SVZ to the center of the olfactory bulb. Netrin-1 secreted from mitral cells attracts DCC- or neogenin-expressing migrating cells, or both, to the olfactory bulb. Slit proteins from the septum inhibit migration out of the RMS into the septum or surrounding tissues by their repulsive activity. Integrins and laminins provide the traction for the motive force, and PSA-N-CAM provides the cellular milieu where the cells can move easily and maintenance of chains. Theshort arrow indicates the differentiation of neural precursors to granule cells; the long arrow marks the periglomerular cells. EPL, External plexiform layer;GCL, granule cell layer; GL, glomerular layer; MCL, mitral cell layer; ONL, olfactory nerve layer.

Article Snippet: The function-blocking anti-β1-integrin rat monoclonal antibody (clone 9EG7) was obtained from PharMingen (San Diego, CA); its specificity has been reported previously ( Lenter et al., 1993 ; Lenter and Vestweber, 1994 ).

Techniques: Migration, Expressing, Activity Assay

Journal: ACS Applied Materials & Interfaces

Article Title: Contact Guidance of Connective Tissue Fibroblasts on Submicrometer Anisotropic Topographical Cues Is Dependent on Tissue of Origin, β1 Integrins, and Tensin-1 Recruitment

doi: 10.1021/acsami.2c22381

Figure Lengend Snippet: Association of α5 and β1 integrin dimers is disrupted on anisotropic repeating groove topographies. Immunofluorescent staining of integrin β1 (green), integrin α5 (red), and nuclei (blue) within HDF and HGF populations following a 24 h timepoint upon the smooth and groove surfaces.

Article Snippet: Fibrillar adhesions were identified using specific antibodies to tensin-1 (NBP1–84129; Novus biologicals, Toronto, ON), integrin β1 (MAB17781; R&D Systems, Minneapolis, MN), and integrin α5 (ab150361; Abcam, Waltham, MA).

Techniques: Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Contact Guidance of Connective Tissue Fibroblasts on Submicrometer Anisotropic Topographical Cues Is Dependent on Tissue of Origin, β1 Integrins, and Tensin-1 Recruitment

doi: 10.1021/acsami.2c22381

Figure Lengend Snippet: Inhibition of integrin αvβ3 and integrin β1 has a differential effect upon HGF and HDF spreading. Immunofluorescent staining of (A) HDF and (B) HGF vinculin (green), F-actin (red), and nuclei (blue) following 1, 3, and 6 h upon a 900 nm periodicity groove surface. The direction of the underlying nanogroove long axis is indicated by white arrows. Scale bar: 20 μm.

Article Snippet: Fibrillar adhesions were identified using specific antibodies to tensin-1 (NBP1–84129; Novus biologicals, Toronto, ON), integrin β1 (MAB17781; R&D Systems, Minneapolis, MN), and integrin α5 (ab150361; Abcam, Waltham, MA).

Techniques: Inhibition, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Contact Guidance of Connective Tissue Fibroblasts on Submicrometer Anisotropic Topographical Cues Is Dependent on Tissue of Origin, β1 Integrins, and Tensin-1 Recruitment

doi: 10.1021/acsami.2c22381

Figure Lengend Snippet: HDF and HGF cell area and circularity are affected by αvβ3 and integrin β1 blocking antibodies The average HDF (A) and HGF (C) area and HDF (B) and HGF (D) circularity in the presence of integrin αvβ3 and β1 blocking antibodies at 1, 3, and 6 h were quantified. Data is expressed as mean ± standard deviation. Data was analyzed using two-way ANOVA, followed by Bonferroni post-tests ( N = 3; * P < 0.05).

Article Snippet: Fibrillar adhesions were identified using specific antibodies to tensin-1 (NBP1–84129; Novus biologicals, Toronto, ON), integrin β1 (MAB17781; R&D Systems, Minneapolis, MN), and integrin α5 (ab150361; Abcam, Waltham, MA).

Techniques: Blocking Assay, Standard Deviation

Journal: ACS Applied Materials & Interfaces

Article Title: Contact Guidance of Connective Tissue Fibroblasts on Submicrometer Anisotropic Topographical Cues Is Dependent on Tissue of Origin, β1 Integrins, and Tensin-1 Recruitment

doi: 10.1021/acsami.2c22381

Figure Lengend Snippet: Independent inhibition of integrin αvβ3 and integrin β1 demonstrates contrast in alignment between HDF and HGF. (A) Immunofluorescent staining of F-actin in HDFs and HGFs at 24 h on a 900 nm periodicity groove surface. (B) Directionality of F-actin expression of integrin αvβ3 and integrin β1 inhibited HDF and (C) HGF following a 24 h timepoint. Mean values are displayed in 10° increments ranging from 0 to 360°. The greatest standard deviation at any point of the distribution is displayed. Data was analyzed using the Komolgorov–Smirnov test between each experimental condition and the control IgG treatment for both HDF and HGF populations ( N = 3; * P < 0.05). The direction of the underlying nanogroove long axis is indicated by white arrows.

Article Snippet: Fibrillar adhesions were identified using specific antibodies to tensin-1 (NBP1–84129; Novus biologicals, Toronto, ON), integrin β1 (MAB17781; R&D Systems, Minneapolis, MN), and integrin α5 (ab150361; Abcam, Waltham, MA).

Techniques: Inhibition, Staining, Expressing, Standard Deviation, Control

Journal: ACS Applied Materials & Interfaces

Article Title: Contact Guidance of Connective Tissue Fibroblasts on Submicrometer Anisotropic Topographical Cues Is Dependent on Tissue of Origin, β1 Integrins, and Tensin-1 Recruitment

doi: 10.1021/acsami.2c22381

Figure Lengend Snippet: Independent inhibition of integrin αvβ3 and integrin β1 demonstrates a functional disparity among (A) HDF and (B) HGF. Immunofluorescent staining of vinculin (green), tensin-1 (red), and nuclei (blue) within HDF and HGF populations, following a 24 h timepoint, upon a 900 nm periodicity nanogroove surface. The direction of the underlying nanogroove long axis is indicated by white arrows.

Article Snippet: Fibrillar adhesions were identified using specific antibodies to tensin-1 (NBP1–84129; Novus biologicals, Toronto, ON), integrin β1 (MAB17781; R&D Systems, Minneapolis, MN), and integrin α5 (ab150361; Abcam, Waltham, MA).

Techniques: Inhibition, Functional Assay, Staining

Journal: ACS Applied Materials & Interfaces

Article Title: Contact Guidance of Connective Tissue Fibroblasts on Submicrometer Anisotropic Topographical Cues Is Dependent on Tissue of Origin, β1 Integrins, and Tensin-1 Recruitment

doi: 10.1021/acsami.2c22381

Figure Lengend Snippet: Inhibition of integrin β1 results in tensin-1 recruitment to αvβ3 peripheral adhesion sites in (A) HDFs, but not (B) HGFs. Immunofluorescent staining of tensin-1 (green), integrin β1 (red), and nuclei (blue) within HDF and HGF populations following a 24 h timepoint. The direction of the underlying nanogroove long axis is indicated by white arrows.

Article Snippet: Fibrillar adhesions were identified using specific antibodies to tensin-1 (NBP1–84129; Novus biologicals, Toronto, ON), integrin β1 (MAB17781; R&D Systems, Minneapolis, MN), and integrin α5 (ab150361; Abcam, Waltham, MA).

Techniques: Inhibition, Staining