Journal: The Journal of Cell Biology

Article Title: Integrin α2β1 Is a Receptor for the Cartilage Matrix Protein Chondroadherin

doi:

Figure Lengend Snippet: β1 integrin-dependent adhesion of chondrocytes to CHAD. Culture dishes (48 well) were coated overnight with chondroadherin (5 μg/ml) and blocked for nonspecific binding with BSA (0.25%). Bovine chondrocytes were allowed to adhere to the dishes for 1 h at 37°C in the presence of various concentrations of a polyclonal antibody against the rat β1 integrin subunit or control IgG. Nonadherent cells were removed by washing, and adhesion was determined by analyzing lysosomal hexosaminidase. The adhesion is expressed as a percentage of the control, and the numbers represent mean of duplicate adhesion from one of three experiments.

Article Snippet: Monoclonal antibody against the human integrin β3 (RUU-PLF12, purified IgG) were purchased from Becton Dickinson (Bedford, MA).

Techniques: Binding Assay, Control

Journal: The Journal of Cell Biology

Article Title: Integrin α2β1 Is a Receptor for the Cartilage Matrix Protein Chondroadherin

doi:

Figure Lengend Snippet: Affinity purification of CHAD-binding cell surface proteins. Bovine chondrocytes were 125 I-labeled and lysed with 1% Triton X-100, 100 μg/ml aprotinin, 2 μg/ml leupeptin, 2 μg/ml pepstatin A, 1 mM PMSF, 1 mM MnCl 2 , and 1 mM MgCl 2 in 10 mM Tris-HCl, pH 7.4. The lysate was passed over control agarose followed by CHAD agarose. Proteins with affinity for CHAD were eluted by EDTA (20 mM), passed over a desalting column (PD-10) equilibrated, and eluted with 0.3 M NaCl, 1% Triton X-100, 0.1% BSA 1 mM CaCl 2 , 1 mM MgCl 2 , 1 mM PMSF, in 50 mM Tris-HCl, pH 7.4. An aliquot of the protein peak was immunoprecipitated with the polyclonal rat β1 integrin antibody. Proteins in the eluate ( E ) and in the immunoprecipitate (β1) were separated by 4–12% SDS-PAGE under reducing ( R ) or nonreducing ( NR ) conditions.

Article Snippet: Monoclonal antibody against the human integrin β3 (RUU-PLF12, purified IgG) were purchased from Becton Dickinson (Bedford, MA).

Techniques: Affinity Purification, Binding Assay, Labeling, Control, Immunoprecipitation, SDS Page

Journal: The Journal of Cell Biology

Article Title: Integrin α2β1 Is a Receptor for the Cartilage Matrix Protein Chondroadherin

doi:

Figure Lengend Snippet: Immunoprecipitation of CHAD-binding integrins from human fibroblasts. 125 I-labeled A549 fibroblasts were lysed with 1% Triton X-100, 100 μg/ml aprotinin, 2 μg/ml leupeptin, 2 μg/ml pepstatin A, 1 mM PMSF, 1 mM MnCl 2 , and 1 mM MgCl 2 in 10 mM Tris-HCl, pH 7.4. The lysate was passed over control agarose followed by CHAD agarose. Proteins with affinity for CHAD were eluted by EDTA (20 mM), passed over a desalting column (PD-10) equilibrated, and eluted with 0.3 M NaCl, 1% Triton X-100, 0.1% BSA, 1 mM CaCl 2 , 1 mM MgCl 2 , 1 mM PMSF in 50 mM Tris-HCl, pH 7.4. Aliquots of the protein peak were immunoprecipitated with monoclonal antibodies against the integrin subunits β1 (P4C10), α1 (TS2/7), α2 (P1E6), α5 (P1D6), and αv (VNR147). The immunoprecipitated proteins were separated by SDS-PAGE (4–12%) under nonreducing conditions and visualized by autoradiography.

Article Snippet: Monoclonal antibody against the human integrin β3 (RUU-PLF12, purified IgG) were purchased from Becton Dickinson (Bedford, MA).

Techniques: Immunoprecipitation, Binding Assay, Labeling, Control, Bioprocessing, SDS Page, Autoradiography

Journal: The Journal of Cell Biology

Article Title: Integrin α2β1 Is a Receptor for the Cartilage Matrix Protein Chondroadherin

doi:

Figure Lengend Snippet: Immunoprecipitation of integrins from human fibroblasts. 125 I-labeled A549 fibroblasts were lysed with 1% Triton X-100, 100 μg/ml aprotinin, 2 μg/ml leupeptin, 2 μg/ml pepstatin A, 1 mM PMSF, 1 mM MnCl 2 , and 1 mM MgCl 2 in 10 mM Tris-HCl, pH 7.4. Aliquots of the lysate were immunoprecipitated with monoclonal antibodies against the integrin subunits β1 (P4C10), β3 (RUU-PLF12), α1 (TS2/7), α2 (P1E6), α3 (P1B5), α5 (P1D6), αv (VNR147), and αvβ5 (P1F6). The immunoprecipitated proteins were separated by SDS-PAGE (4–12%) under nonreducing conditions and visualized by autoradiography.

Article Snippet: Monoclonal antibody against the human integrin β3 (RUU-PLF12, purified IgG) were purchased from Becton Dickinson (Bedford, MA).

Techniques: Immunoprecipitation, Labeling, Bioprocessing, SDS Page, Autoradiography

Journal: The Journal of Cell Biology

Article Title: Integrin α2β1 Is a Receptor for the Cartilage Matrix Protein Chondroadherin

doi:

Figure Lengend Snippet: Inhibition of fibroblast adhesion to CHAD by integrin antibodies. Culture dishes (48 well) were coated with CHAD (5 μg/ml) and blocked for nonspecific binding with BSA (0.25%). Human A549 fibroblasts were allowed to adhere to the dishes for 1 h at 37°C in the presence of monoclonal antibodies against the human integrin subunits β1 (P4C10), β3 (RUU-PLF12), α2 (Gi9), α3 (P1B5), α5 (P1D6), αv (VNR147), αvβ3 (LM609), and αvβ5 (P1F6). Nonadherent cells were removed by washing, and adhesion was determined by analyzing lysosomal hexosaminidase. The adhesion is expressed as a percentage of the control, and the numbers represent the mean of duplicate adhesion from three individual experiments ±SD. * P < 0.05; ** P < 0.01; P β1 = 0.002; P α2 = 0.011; P α3 = 0.021.

Article Snippet: Monoclonal antibody against the human integrin β3 (RUU-PLF12, purified IgG) were purchased from Becton Dickinson (Bedford, MA).

Techniques: Inhibition, Binding Assay, Bioprocessing, Control

Journal: The Journal of Cell Biology

Article Title: Integrin α2β1 Is a Receptor for the Cartilage Matrix Protein Chondroadherin

doi:

Figure Lengend Snippet: Inhibition of human chondrocyte adhesion to CHAD by α2 integrin antibodies. Culture dishes (48 well) were coated with CHAD (5 μg/ml) and blocked for nonspecific binding with BSA (0.25%). Human chondrocytes were allowed to adhere to the dishes for 1 h at 37°C in the presence of various concentrations of the monoclonal antibody against the human integrin subunit α2 (Gi9). Nonadherent cells were removed by washing, and adhesion was determined by analyzing lysosomal hexosaminidase. The adhesion is expressed as a percentage of the control, and the numbers represent the mean adhesion ±SD from three wells in one of two experiments.

Article Snippet: Monoclonal antibody against the human integrin β3 (RUU-PLF12, purified IgG) were purchased from Becton Dickinson (Bedford, MA).

Techniques: Inhibition, Binding Assay, Control

Journal: The Journal of Cell Biology

Article Title: Integrin α2β1 Is a Receptor for the Cartilage Matrix Protein Chondroadherin

doi:

Figure Lengend Snippet: Inhibition of adhesion of T47D cells to CHAD ( a ) and to collagen type II (CII; b ) by various α2 antibodies. Culture dishes (48 wells) were coated with 5 μg/ml of CHAD or CII and blocked for nonspecific binding with BSA (0.25%). T47D cells were allowed to adhere to the dishes for 1 h at 37°C in the absence or in the presence of monoclonal antibodies against the integrin subunits β1 (P4C10), β3 (RUU-PLF12), or various α2 antibodies. Nonadherent cells were removed by washing, and adhesion was determined by analyzing lysosomal hexosaminidase. The adhesion is expressed as a percentage of the control, and the numbers represent the mean of duplicate adhesion from three individual experiments ±SD. * P < 0.05; ** P < 0.01; ( a ) P β1 = 0.004; P Gi9 = 0.006; P Gi19 = 0.026. ( b ) P β1 = 0.000; P P1E6 = 0.001; P P1H5 = 0.001; P Gi9 = 0.000; P Gi26 = 0.047.

Article Snippet: Monoclonal antibody against the human integrin β3 (RUU-PLF12, purified IgG) were purchased from Becton Dickinson (Bedford, MA).

Techniques: Inhibition, Binding Assay, Bioprocessing, Control

Journal: World Journal of Hepatology

Article Title: Non-invasively differentiate non-alcoholic steatohepatitis by visualizing hepatic integrin αvβ3 expression with a targeted molecular imaging modality

doi: 10.4254/wjh.v16.i11.1290

Figure Lengend Snippet: Expression of integrin αvβ3 in cultured hepatocytes. Human LO2 hepatocytes were cultured with the medium containing 100 μmol/L palmitate acid and 200 μmol/L oleic acid (FFA) for 24 hours, and the cells cultured in the medium without FFA served as the control. A: Representative micrographs of the control and FFA-cultured hepatocytes after stained with Oil-Red O. Images were taken at original magnification (200 ×), scale bars = 20 μm; B and C: Representative fluorescent images of the control (B) and FFA-cultured (C) hepatocytes after separately stained with integrin αv and β3 subunits antibody (green color) and counterstained with albumin antibody (red color). 6-diamidino-2-phenylindole was used for nuclei staining. The merged images show the yellow color area by overlaying images of the counterstaining. Images were taken at original magnification (400 ×), scale bars = 100 μm; D: Comparison of the message RNA levels of integrin αv and β3 subunits in the control and FFA-cultured hepatocytes. The message RNA levels of integrin αv and β3 subunit were determined by quantitative real-time polymerase chain reaction analysis; E and F: Comparison of the protein amounts of integrin αv and β3 subunits in the control and FFA-cultured hepatocytes. The protein amounts of integrin αv and β3 subunits were analyzed by western-blot assay, and β-Tublin was used as the reference. All experiments were undertaken in triplicates. In all panels, data are expressed in means ± SD. FFA: Oleic acid; DAPI: 6-diamidino-2-phenylindole.

Article Snippet: Secondly, after being fixed with 4% paraformaldehyde and permeabilized with phosphate-buffered saline (PBS) containing 0.1% Triton X-100 and 0.1 mg/mL RNase A when appropriate, the control and FFA-cultured hepatocytes were incubated with primary antibodies against integrin αv subunit (1:1000, Abmart, T56887) or integrin β3 subunit (1:1000, Abmart, M015908), and albumin (1:1000, Servicebio, GB122080) at 4 °C overnight.

Techniques: Expressing, Cell Culture, Control, Staining, Comparison, Real-time Polymerase Chain Reaction, Western Blot

Journal: World Journal of Hepatology

Article Title: Non-invasively differentiate non-alcoholic steatohepatitis by visualizing hepatic integrin αvβ3 expression with a targeted molecular imaging modality

doi: 10.4254/wjh.v16.i11.1290

Figure Lengend Snippet: Expression of integrin αvβ3 in livers of rabbits with non-alcoholic fatty liver disease. Non-alcoholic fatty liver disease was induced in rabbits by high-fat diet (HFD) for 2, 6, and 8 months (referred to as HFD-2M, 6M and 8M), and rabbits fed with regular diet for 8 months served as the control group ( n = 6 per group). A: Representative micrographs of hepatic histology stained with hematoxylin-eosin staining (200 ×), Sirius red (100 ×), and immunohistochemistry for integrin β3 subunit (400 ×). In immunohistochemistry staining images, the brown areas indicated integrin β3 subunit positive staining, scale bars = 50 μm; B: Comparison of non-alcoholic fatty liver disease activity score in the control and HFD-fed rabbits; C: Comparison of the percentage of integrin β3 subunit positive-staining area in liver sections of the control and HFD-fed rabbits. For semi-quantitative analysis of hepatic integrin αvβ3 expression level, 10 fields were randomly selected and recorded from each section stained with immunohistochemistry for integrin β3 subunit. Then integrin β3 subunit positive-staining area was measured, and the percentages of the positive-staining area in liver sections were compared. In all panels, data are expressed in means ± SD. IHC: Immunohistochemistry; HE: Hematoxylin-eosin staining; CTRL: Normal control group; M: Month; NS: Not significant; NAFLD: Non-alcoholic fatty liver disease.

Article Snippet: Secondly, after being fixed with 4% paraformaldehyde and permeabilized with phosphate-buffered saline (PBS) containing 0.1% Triton X-100 and 0.1 mg/mL RNase A when appropriate, the control and FFA-cultured hepatocytes were incubated with primary antibodies against integrin αv subunit (1:1000, Abmart, T56887) or integrin β3 subunit (1:1000, Abmart, M015908), and albumin (1:1000, Servicebio, GB122080) at 4 °C overnight.

Techniques: Expressing, Control, Staining, Immunohistochemistry, Comparison, Activity Assay

Journal: World Journal of Hepatology

Article Title: Non-invasively differentiate non-alcoholic steatohepatitis by visualizing hepatic integrin αvβ3 expression with a targeted molecular imaging modality

doi: 10.4254/wjh.v16.i11.1290

Figure Lengend Snippet: Expression of integrin αvβ3 in livers of rats with non-alcoholic fatty liver disease. Non-alcoholic fatty liver disease was induced in rats by high-fat high-carbohydrate diet (HFCD) for 2, 4, and 6 months (referred to as HFCD-2M, 4M and 6M), and rats fed with regular diet for 6 months served as the control group ( n = 6 per group). A: Representative micrographs of hepatic histology stained with H&E (200 ×), Masson (100 ×), and Oil-red O (200 ×), scale bars = 50 μm; B: Comparison of non-alcoholic fatty liver disease activity score in the control and HFCD-fed rats; C: Comparison of hepatic integrin αvβ3 message RNA level in the control and HFCD-fed rats. Hepatic message RNA levels of integrin αv and β3 subunits were respectively determined by quantitative real-time polymerase chain reaction analysis; D-F: Comparison of the protein amounts of integrin αvβ3 in livers of the control and HFCD-fed rats. The protein amounts of integrin αv and β3 subunits were analyzed by western-blot assay, and β-Tublin was used as the reference. In all panels, data are expressed in means ± SD. HE: Hematoxylin-eosin staining; CTRL: Normal control group; M: Month; NS: Not significant; NAFLD: Non-alcoholic fatty liver disease.

Article Snippet: Secondly, after being fixed with 4% paraformaldehyde and permeabilized with phosphate-buffered saline (PBS) containing 0.1% Triton X-100 and 0.1 mg/mL RNase A when appropriate, the control and FFA-cultured hepatocytes were incubated with primary antibodies against integrin αv subunit (1:1000, Abmart, T56887) or integrin β3 subunit (1:1000, Abmart, M015908), and albumin (1:1000, Servicebio, GB122080) at 4 °C overnight.

Techniques: Expressing, Control, Staining, Comparison, Activity Assay, Real-time Polymerase Chain Reaction, Western Blot

Journal: World Journal of Hepatology

Article Title: Non-invasively differentiate non-alcoholic steatohepatitis by visualizing hepatic integrin αvβ3 expression with a targeted molecular imaging modality

doi: 10.4254/wjh.v16.i11.1290

Figure Lengend Snippet: Immunofluorescent co-localization of integrin αvβ3 and the markers of various hepatic cells including α-smooth muscle actin, cluster of differentiation 31, cluster of differentiation 68 or cluster of differentiation 163 in livers of the control and high-fat high-carbohydrate diet-fed rats. Representative fluorescent images of integrin β3 subunit (green color) and albumin, α-smooth muscle actin, cluster of differentiation (CD) 31, CD68 and CD163 (red color) in liver sections, which were separately stained with specific first antibodies and visualized by second antibodies, and counterstained with 6-diamidino-2-phenylindole for nuclei staining. The merged images show the yellow color area by overlaying images of the counterstaining, and amplified images corresponding to the indicated areas in boxes are present. Images were recorded at original magnification (400 ×), scale bars = 100 μm. A: Control; B: High-fat high-carbohydrate diet (HFCD)-2M; C: HFCD-4M; D: HFCD-6M rats; E: Comparison of the percentage of integrin β3 subunit positive-staining area in liver sections of the control and HFCD-fed rats. Ten fields were randomly selected and recorded from each section. Then integrin β3 positive-staining area was measured, and the percentages of the positive-staining area in liver sections were compared; F: Comparison of the ratio of the overlapped yellow area to integrin β3 subunit positive-staining green area in liver sections of HFCD-6M rats. Ten fields were randomly selected and recorded from each section. Then integrin β3 subunit positive-staining area and the area of integrin β3 subunit positive-staining overlapped with hepatic cellular markers positive-staining were respectively measured, and the ratios were compared. In all panels, data are expressed in means ± SD. DAPI: 6-diamidino-2-phenylindole; CTRL: Normal control group; M: Month; CD: Cluster of differentiation; αSMA: α-smooth muscle actin.

Article Snippet: Secondly, after being fixed with 4% paraformaldehyde and permeabilized with phosphate-buffered saline (PBS) containing 0.1% Triton X-100 and 0.1 mg/mL RNase A when appropriate, the control and FFA-cultured hepatocytes were incubated with primary antibodies against integrin αv subunit (1:1000, Abmart, T56887) or integrin β3 subunit (1:1000, Abmart, M015908), and albumin (1:1000, Servicebio, GB122080) at 4 °C overnight.

Techniques: Control, Staining, Amplification, Comparison

Journal: The Journal of Biological Chemistry

Article Title: The dipeptide prolyl-hydroxyproline promotes cellular homeostasis and lamellipodia-driven motility via active β1-integrin in adult tendon cells

doi: 10.1016/j.jbc.2021.100819

Figure Lengend Snippet: Engagement of β1-integrin and ERK signaling in Pro-Hyp-mediated cell migration. A , left panels , immunofluorescence staining for β1-integrin (clone 9EG7, which recognizes an extracellular epitope of ligand-inducible active β1 : green )/DAPI ( blue ) and F-actin ( red )/DAPI ( blue ). Adult tenocytes were treated with 500 μg/ml Pro-Hyp for 6 h. The scale bars represent 20 μm. Right panels , the numbers of active β1-integrin–containing focal contacts and the thickness of F-actin filaments when cells were treated with 500 μg/ml Pro-Hyp for 6 h. Error bars represent the standard deviation (n = 4; four separate culture experiments). ∗∗ p < 0.01; ∗∗∗ p < 0.001. B , effect of MEK1/2 inhibitor PD98059 or α5β1-integrin antagonist ATN-161 on Pro-Hyp–mediated cell migration in adult tenocytes using Boyden chamber. Cell migration activity is shown relative to the control value of 100% (cells treated with 200 μg/ml Pro-Hyp). Error bars represent the standard deviation (n = 5). ∗∗∗ p < 0.001 (in post hoc analysis). C , effect of ATN-161 on uptake of stable isotopically labeled Pro-Hyp (SI-Pro-Hyp; 200 μg/ml) in adult tenocytes for 60 min at 37 °C. Error bars represent the standard deviation (n = 8). ∗∗∗ p < 0.001. DAPI, 4′,6-diamidino-2-phenylindole; ERK, extracellular signal–regulated kinase; MEK1/2, MAPK kinase 1/2; Pro-Hyp, prolyl-4-hydroxyproline.

Article Snippet: The following antibodies were used: rabbit polyclonal antibody (pAb) against mouse type I collagen (Chemicon: this antibody shows less than 0.1% reactivity with mouse collagen types II and IV in addition to 1.0% reactivity with mouse collagen type III); rabbit pAb against mouse fibronectin (Chemicon: this antibody has crossreactivity to bovine fibronectin and shows less than 0.1% reactivity with mouse laminin and collagen types I, III, and IV by radioimmunoassay); rabbit pAb against the C-telopeptide of the α1 chain of type I collagen (LF68; provided by Dr Larry Fisher, the National Institutes of Health, USA); rabbit pAb (Abcam) and goat pAb (Thermo Fisher) against human type V collagen; mouse mAb against phospho-ERK1/2 (pT202/pY204) and rabbit pAb against total Erk (Cell Signaling); rat mAb against mouse integrin β1 (clone 9EG7) , hamster mAbs against mouse integrin β1 (clone Ha2/5), mouse integrin α1 (clone Ha31/8), and mouse integrin α2 (clone Ha1/29); rat mAbs against mouse integrin α5 (clone 5H10-27), mouse integrin α6 (clone GoH3), and mouse integrin αv (clone RMV-7) and hamster mAb against mouse integrin β3 (clone 2C9.G2) (all from Pharmingen); α11 (rabbit pAb; provided by Dr Donald Gullberg, University of Bergen, Norway); rabbit pAb against human calnexin (sc-11397; Santa Cruz); rabbit pAb against human Histone H1.0 (GeneTex); mouse mAb against human vimentin (clone VIM-13.2; Sigma); mouse mAb against α-tubulin (clone B-5-1-2; Sigma); and mouse mAb against rabbit GAPDH (clone 3H12; MBL).

Techniques: Migration, Immunofluorescence, Staining, Standard Deviation, Activity Assay, Labeling

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: Integrin expression profiles are altered after OC-2 cell differentiation . (A) Western blot analysis of myosin VI and myosin VIIa levels in OC-2 cells grown at 33°C and 39°C. Both myosin VI and VIIa levels were increased significantly in cells grown at 39°C cells in comparison with those incubated at 33°C. Bar graphs represent densitometric results of mean relative values of myosin VI and myosin VIIa levels ± s.e.m. HSC-70 provided the loading control. (B) Undifferentiated (33°C) and differentiated (39°C) OC-2 cells were analysed by FACS for the integrin subunits α6, β1, αv and β3. Levels of β1-, αv- and β3-integrin subunits were increased significantly in differentiated OC-2 cells when compared with undifferentiated OC-2 cells. α6-integrin surface expressions levels did not change between the two cell phenotypes. Bar graph represents mean fluorescence units of the various integrin subunits ± s.e.m.; n = 3 independent experiments. White bars = cells at 33°C, black bars = cells at 39°C cells; nd = no significant difference, * P < 0.05, ** P < 0.01.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing, Cell Differentiation, Western Blot, Incubation, Fluorescence

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: Integrin expression profiles in the Organ of Corti . RT-PCR of RNA isolated from organ of Corti of adult mice. Bands at the predicted molecular weight are visible for integrin β1, β3, αv, α6 and β-actin but not for integrin β4. cDNA was omitted for the negative control.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Molecular Weight, Negative Control

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: αv-integrin expression profiles change during differentiation . Expression of (A) α6-integrin and (B) αv-integrin were examined by FACS at days 2, 6, 10 and 14 during the differentiation process. The differentiating cells ('dif' red line) were compared to undifferentiated ('33' green line) and fully differentiated cells ('39' red line). α6-integrin expression profiles did not change throughout the time course. αv-integrin expression profiles show an increase at day 6 and are at comparable levels with fully differentiated OC-2 cells at day 14 with an apparent even higher level at day 10.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: β1 and β3-integrin expression profiles change during differentiation . Surface expression of (A) β1-integrin and (B) β3-integrin was examined by FACS at days 2, 6, 10 and 14 of the differentiation process The differentiating cells ('dif' red line) were compared to undifferentiated ('33' green line) and fully differentiated cells ('39' red line). Increase of β1-integrin expression profile is visible from day 2 and it reaches the level of differentiated cells by day 10. β3-integrin expression which is undetectable above the Ig signal up to day 2, becomes visible at day 6 and gradually reaches the differentiated levels at day 14.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: β3-integrin is a marker for OC-2 cell differentiation . (A) Western blot analysis of myosin VIIa expression in 39°C OC-2 cells untreated or after treatment with either scrambled (Scr), β1-, α6-, αv- and β3-integrin siRNA. Treatment with all siRNAs reduced myosin VIIa levels significantly when compared with untreated OC-2 cells or OC-2 cells treated with scrambled siRNA. (B) Undifferentiated OC-2 cells transduced with human β3-integrin (+hβ3) had similar levels of surface β3-integrin to differentiated (39) OC-2 cells. Furthermore, analysis of the expression of myosin VIIa showed it was increased significantly in OC-2 cells transduced with human β3-integrin (+hβ3) cells compared with undifferentiated (33) cells. Bar graphs represent densitometric results of mean relative values of myosin VIIa levels ± s.e.m. HSC-70 was used as a loading control. * P < 0.05, ** P < 0.01. n = 3 individual experiments.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Marker, Cell Differentiation, Western Blot, Expressing, Transduction

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: Effect of β3-integrin-siRNA treatment on integrin surface expression . Differentiated OC2 cells were treated with β3-siRNA and assayed by FACS for expression of surface integrins. The data is compared to integrin expression levels in normal differentiated cells (39°), differentiated cells exposed to scrambled siRNA sequences (siRNAβ3) and undifferentiated cells (33°). Exposure to β3-integrin-siRNA resulted in a significant reduction of the surface expression of β3, β1- and αv- to levels similar to those of undifferentiated cells. There was no significant effect of scrambled siRNA sequences on any integrin expression and β3-integrin-siRNA did not significantly affect α6-integrin providing further evidence for the specificity of the siRNA inhibition.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing, Inhibition

Journal: BMC Cell Biology

Article Title: β3-integrin is required for differentiation in OC-2 cells derived from mammalian embryonic inner ear

doi: 10.1186/1471-2121-13-5

Figure Lengend Snippet: Graph of changes in expression levels of integrin subunits and myosin VIIa during OC-2 cell differentiation . Schematic time course graph showing fluorescence values of αv-, α6-, β1- and β3- integrin subunit surface expression measured by FACS and chemiluminescence values of the hair cell marker myosin VIIa in OC-2 cells measured by Western blot, over the 14 day differentiation process.

Article Snippet: Biotinylated β3-integrin antibody was purchased from Pharmingen (Bedford, UK). αv-integrin antibody was purchased from Chemicon International (Harrow, UK). α6-integrin antibody was purchased from ABD Serotec (MorphoSys Ltd, Kidlington, UK). β4-integrin antibody was purchased from AbCam (Cambridge, UK).

Techniques: Expressing, Cell Differentiation, Fluorescence, Marker, Western Blot

Journal:

Article Title: Laminin-6 assembles into multimolecular fibrillar complexes with perlecan and participates in mechanical-signal transduction via a dystroglycan-dependent, integrin-independent mechanism

doi: 10.1242/jcs.02395

Figure Lengend Snippet: Fig. 3

Article Snippet: Mouse monoclonal antibodies against the γ2 and β3 subunits of rat laminin 5 were originally provided by Desmos (San Diego, CA).

Techniques: