Journal: Cell metabolism

Article Title: Changes in integrin expression during adipocyte differentiation.

doi: 10.1016/j.cmet.2005.08.006

Figure Lengend Snippet: Figure 2. Expression switch of integrin α subunits during differentiation of preadipocytes

Article Snippet: The C/EBPα (14AA), Myc (C-19), integrin α5 (H-104), and α6 (H-87) polyclonal antibodies were purchased from Santa Cruz Inc.

Techniques: Expressing

Journal: Cell metabolism

Article Title: Changes in integrin expression during adipocyte differentiation.

doi: 10.1016/j.cmet.2005.08.006

Figure Lengend Snippet: Figure 5. Clustering of growth-arrested preadipocytes on Matrigel depends on integrin α6

Article Snippet: The C/EBPα (14AA), Myc (C-19), integrin α5 (H-104), and α6 (H-87) polyclonal antibodies were purchased from Santa Cruz Inc.

Techniques:

Journal: PLoS Pathogens

Article Title: Serum bridging molecules drive candidal invasion of human but not mouse endothelial cells

doi: 10.1371/journal.ppat.1010681

Figure Lengend Snippet: (A) Western blot showing that endothelial cell gC1qR binds to C . glabrata coated with fresh serum. Results are representative of 3 independent experiments. (B-E) Effects of inhibiting gC1qR function with siRNA knockdown (B and C) and specific monoclonal antibodies (D and E) on the endocytosis (B and D) and cell-association (C and E) of C . glabrata coated with fresh serum. Antibody 74.5.2 recognizes the high molecular weight kininogen binding site in the C-terminus of the gC1qR and antibody 60.11 is directed against the C1q binding site in the N-terminus of the gC1qR. Results shown in (B-E) are the mean ± SD of 3 experiments each performed in triplicate. Heat inact., heat inactivated serum; orgs/HPF, organisms per high-power field; ns, not significant, * P < 0.05, ** P < 0.01 by Student’s t-test (B and C) or ANOVA with the Dunnett’s test for multiple comparisons (D and E).

Article Snippet: The endothelial cells were transfected with gC1qR siRNA (Santa Cruz Biotechnology, Cat. # sc-42880), integrin α5 siRNA (Santa Cruz Biotechnology, Cat. # sc-29372), integrin αv siRNA (Santa Cruz Biotechnology, Cat. # sc-29373), or scrambled control siRNA (Qiagen, Cat. # 1027281) using Lipofectamine 2000 (Thermo Fisher Scientific) following the manufacturer’s instructions.

Techniques: Western Blot, Knockdown, Bioprocessing, High Molecular Weight, Binding Assay

Journal: PLoS Pathogens

Article Title: Serum bridging molecules drive candidal invasion of human but not mouse endothelial cells

doi: 10.1371/journal.ppat.1010681

Figure Lengend Snippet: (A-F) Effects of inhibiting αv integrin function with siRNA knockdown (A and B or)specific monoclonal antibodies (C-F) and on the endocytosis (A, C, D) and cell-association (B, E, F) of serum-coated C . glabrata . (G and H) Inhibition of gC1qR (with monoclonal antibody 74.5.2) and αv integrins has an additive effect on decreasing the endocytosis (G) but not cell-association of serum-coated C . glabrata (H). Results are the mean ± SD of 3 experiments, each performed in triplicate. Orgs/HPF, organisms per high power field; ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by ANOVA with the Dunnett’s test for multiple comparisons (A, B, G, H) or the Student’s t-test (C-F).

Article Snippet: The endothelial cells were transfected with gC1qR siRNA (Santa Cruz Biotechnology, Cat. # sc-42880), integrin α5 siRNA (Santa Cruz Biotechnology, Cat. # sc-29372), integrin αv siRNA (Santa Cruz Biotechnology, Cat. # sc-29373), or scrambled control siRNA (Qiagen, Cat. # 1027281) using Lipofectamine 2000 (Thermo Fisher Scientific) following the manufacturer’s instructions.

Techniques: Knockdown, Bioprocessing, Inhibition

Journal: PLoS Pathogens

Article Title: Serum bridging molecules drive candidal invasion of human but not mouse endothelial cells

doi: 10.1371/journal.ppat.1010681

Figure Lengend Snippet: (A and B) Endocytosis of C . glabrata coated with either human or mouse serum by the indicated endothelial cells after 45 min (A) and 180 min (B). (C) Endocytosis of C . glabrata coated with fresh human serum by mouse liver endothelial cells expressing human gC1qR, integrin αv, or integrin β5. Data are the mean ± SD of 3 experiments each performed in triplicate. HUVEC, human umbilical vein endothelial cell; orgs/HPF, organisms per high power field; ns, not significant; ** P < 0.01, **** P < 0.0001. *** P < 0.001, **** P < 0.0001 by ANOVA with the Dunnett’s test for multiple comparisons.

Article Snippet: The endothelial cells were transfected with gC1qR siRNA (Santa Cruz Biotechnology, Cat. # sc-42880), integrin α5 siRNA (Santa Cruz Biotechnology, Cat. # sc-29372), integrin αv siRNA (Santa Cruz Biotechnology, Cat. # sc-29373), or scrambled control siRNA (Qiagen, Cat. # 1027281) using Lipofectamine 2000 (Thermo Fisher Scientific) following the manufacturer’s instructions.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Integrin VLA-5 and FAK are Good Targets to Improve Treatment Response in the Philadelphia Chromosome Positive Acute Lymphoblastic Leukemia

doi: 10.3389/fonc.2014.00112

Figure Lengend Snippet: The expression level of integrin α5 subunit in Ph+ leukemia cell line SUP-B15 is significantly increased after serum starvation . (A) The percentages of cells expressing integrin α4, α5, and β1 subunits before and after serum starvation for 7 h detected by flow cytometry. (B) The mean fluorescent intensity (MFI) of integrin α4, α5, and β1 subunits from whole alive cell population before and after serum starvation for 7 h detected by flow cytometry.

Article Snippet: Human integrin α5 shRNA lentiviral particles (Catalog # sc-29372-V, Santa Cruz) were thawed at room temperature and added to leukemia cell suspension in 15 ml conical tubes and were spun at 800 × g (2500 rpm) for 90 min at 37°C.

Techniques: Expressing, Flow Cytometry

Journal: Frontiers in Oncology

Article Title: Integrin VLA-5 and FAK are Good Targets to Improve Treatment Response in the Philadelphia Chromosome Positive Acute Lymphoblastic Leukemia

doi: 10.3389/fonc.2014.00112

Figure Lengend Snippet: Integrin α5 subunit antibody inhibits the adhesion of Ph+ leukemia cells to human fibronectin and enhances the killing of imatinib . (A) Cell adhesion assay showed that α5 subunit inhibitory antibody (CD49e, clone IIA1 BD Biosciences) was the only tested integrin antibody that significantly inhibited the adhesion of Ph+ leukemia cells to fibronectin with adhesion percentage of 6.6 ± 3.8% compared with control IgG of 44.8 ± 7.9%, p < 0.01. (B) . Annexin-V plus PI by flow cytometry. (i) P2 gate identifies the CD38 positive cell population representing SUP-B15 Ph+ leukemia cells. (ii) Ph+ leukemia cells grown on HS-5 stromal cells. (iii) Ph+ leukemia cells treated with 10 μM imatinib (IM). (iv) SUP-B15 cultured with stromal cells treated with 10 μM imatinib. (v) SUP-B15 cultured with stromal cells treated with 10 μM imatinib and 10 μg/ml anti-α5 Ab. (C) Quantitative analysis for apoptosis rate of different conditions. *After culture on HS-5 cells for 24 h, the apoptosis rate of leukemia cells SUP-B15 decreased from 29.4 ± 2.3 to 16.7 ± 3%, p < 0.05. # When inhibitory antibody to integrin α5 was combined with imatinib to treat Ph+ leukemia cells cultured on stromal cells, the apoptosis rate 38.0 ± 8.0% was significantly increased compared with imatinib by itself 25.7 ± 3.3%, p < 0.05.

Article Snippet: Human integrin α5 shRNA lentiviral particles (Catalog # sc-29372-V, Santa Cruz) were thawed at room temperature and added to leukemia cell suspension in 15 ml conical tubes and were spun at 800 × g (2500 rpm) for 90 min at 37°C.

Techniques: Cell Adhesion Assay, Control, Flow Cytometry, Cell Culture

Journal: Frontiers in Oncology

Article Title: Integrin VLA-5 and FAK are Good Targets to Improve Treatment Response in the Philadelphia Chromosome Positive Acute Lymphoblastic Leukemia

doi: 10.3389/fonc.2014.00112

Figure Lengend Snippet: Blocking integrin α5 affects the engraftment of Ph+ leukemia cells in immunodeficient mice . (A) The incubation for 1 h of Ph+ leukemia cells with disintegrin, a peptide inhibitor of integrins, impaired the engraftment of leukemia in NSG mice. Representative figures of n = 2 showed bioluminescence imaging from day 7 to 28 after inoculation of leukemia cells. (B) Animal total body bioluminescence was measured using the Xenogen IVIS Imaging System 200 Series with total imaging time of 2 min and compared with control animals that received cells that were not treated with Echistatin. (C) Anti-integrin α5 inhibitory antibodies clone IIA1 (BD Biosciences) and clone P1D6 (Millipore) decreased the engraftment of Ph+ leukemia cells in the bone marrow of NOD/SCID mice ( n = 2).

Article Snippet: Human integrin α5 shRNA lentiviral particles (Catalog # sc-29372-V, Santa Cruz) were thawed at room temperature and added to leukemia cell suspension in 15 ml conical tubes and were spun at 800 × g (2500 rpm) for 90 min at 37°C.

Techniques: Blocking Assay, Incubation, Imaging, Control

Journal: Frontiers in Oncology

Article Title: Integrin VLA-5 and FAK are Good Targets to Improve Treatment Response in the Philadelphia Chromosome Positive Acute Lymphoblastic Leukemia

doi: 10.3389/fonc.2014.00112

Figure Lengend Snippet: Knocking down integrin α5 delayed the engraftment of Ph+ leukemia cells in immunodeficient NOD/SCID mice . (A) Real-time QPCR assay showed that we had successfully knocked down the expression level of the integrin α5 knocking in clone 10. The cycle threshold (Ct) of the target gene (integrin α5) was normalized to the chosen reference gene GAPHD. Relative quantification, R = 2 −(ΔCt sample − ΔCt control) . Result shows the fold change normalized to SUP-LUC2 cells. (B) Western blot showed a reduced protein expression level of integrin α5 in clone 10. (C) Flow cytometry showed reduced mean fluorescent intensity of integrin α5 in clone 10. (D) Representative figures showed bioluminescent imaging at 2 weeks, and 5–8 weeks post leukemia cells inoculation. (E) Animal total body bioluminescence was quantified to compare with control. **The levels of bioluminescence became most significantly different 2 months post inoculation between the α5 knock-down group and control group [mean (SD) radiance vs. control, 5.3 (0.1) vs. 2.1 (0.2) × 10 6 p/s/cm 2 /sr, p < 0.01 at 2 months, n = 3.].

Article Snippet: Human integrin α5 shRNA lentiviral particles (Catalog # sc-29372-V, Santa Cruz) were thawed at room temperature and added to leukemia cell suspension in 15 ml conical tubes and were spun at 800 × g (2500 rpm) for 90 min at 37°C.

Techniques: Expressing, Quantitative Proteomics, Control, Western Blot, Flow Cytometry, Imaging, Knockdown

Journal: BioMed Research International

Article Title: Participation of Integrin α 5 β 1 in the Fibronectin-Mediated Adherence of Enteroaggregative Escherichia coli to Intestinal Cells

doi: 10.1155/2014/781246

Figure Lengend Snippet: Integrin α 5 expression knockdown reduces EAEC strain 042 fibronectin-mediated binding to epithelial cells. HEp-2 cells nontransfected and transfected with scrambled or integrin α 5 shRNA were preincubated with DMEM only (control) or medium supplemented with fibronectin (Fn) and then infected with EAEC strain 042. Numbers of adherent bacteria were determined by colony forming unit counts (CFU). *Significantly different compared to control treatment ( P < 0.05).

Article Snippet: Subconfluent cultures (~40–50%) of HEp-2 cells grown in 6-well plates were transfected with small hairpin RNA (shRNA) for integrin α 5 (sc-29372-SH; Santa Cruz Biotechnology) following the manufacturer's instructions (Santa Cruz Biotechnology).

Techniques: Expressing, Knockdown, Binding Assay, Transfection, shRNA, Control, Infection, Bacteria

Journal: International journal of oncology

Article Title: Syntaxins 3 and 4 mediate vesicular trafficking of α5β1 and α3β1 integrins and cancer cell migration.

doi: 10.3892/ijo.2011.1101

Figure Lengend Snippet: Figure 2. siRNA knockdown of syntaxins (STX) 3 and 4 diminishes cell surface expression of α5 integrin. (A) HeLa cells were transfected with a non- targeting control siRNA or a siRNA to syntaxins 2, 3 or 4. Seventy-two hours after transfection, whole cell lysates of the cells were immunoblotted with an antibody to syntaxins 2, 3 or 4. The same membranes were reprobed with an antibody to β-actin as a loading control. (B) The transfected cells were stained with an antibody to α5 integrin without permeabilization. Representative confocal images of 3 independent experiments are shown. In each experiment, images were collected at the same pinhole, detector gain and laser intensity. Scale bar, 50 µm. (C) 30 µg of whole cell lysates of the transfected cells were immunoblotted with an antibody to α5 integrin.

Article Snippet: The membranes were blotted with a rabbit pAb to syntaxin 2, a rabbit pAb to syntaxin 3, a mouse mAb to syntaxin 4 (Sigma-Aldrich Co.), a rabbit pAb to α5 integrin (Santa Cruz Biotechnology), a mouse mAb to β-actin (Sigma-Aldrich Co.), and HRP-conjugated secondary antibodies.

Techniques: Knockdown, Expressing, Transfection, Control, Staining

Journal: International journal of oncology

Article Title: Syntaxins 3 and 4 mediate vesicular trafficking of α5β1 and α3β1 integrins and cancer cell migration.

doi: 10.3892/ijo.2011.1101

Figure Lengend Snippet: Figure 3. Accumulation of α5 and α3 integrins in cytoplasmic vesicles in syntaxins 3 and 4 knockdown cells. Seventy-two hours after transfection with the control, syntaxins 2, 3 or 4 siRNAs, HeLa cells were permeabilized and stained with an antibody to α5 integrin (top row) or α3 integrin (bottom row). Representative confocal images are shown. Scale bar, 20 µm.

Article Snippet: The membranes were blotted with a rabbit pAb to syntaxin 2, a rabbit pAb to syntaxin 3, a mouse mAb to syntaxin 4 (Sigma-Aldrich Co.), a rabbit pAb to α5 integrin (Santa Cruz Biotechnology), a mouse mAb to β-actin (Sigma-Aldrich Co.), and HRP-conjugated secondary antibodies.

Techniques: Knockdown, Transfection, Control, Staining

Journal: Oncology Reports

Article Title: Overexpression of SSAT by DENSPM treatment induces cell detachment and apoptosis in glioblastoma

doi: 10.3892/or.2011.1592

Figure Lengend Snippet: Reduced expression of anti-apoptosis related proteins AKT, mTOR and the adhesion-related proteins integrin α5, integrin β1 in PCMV-SSAT-LN229 cells.

Article Snippet: Incubation with primary polyclonal antibodies against integrin α5 (BD Biosciences, San Jose, CA), integrin β1 (BD Biosciences), AKT, mTOR (Cell Signaling Technology, Danvers, MA), SSAT (Santa Cruz Biotechnology, Santa Cruz, CA) and actin (Sigma, St. Louis, MO) were performed at a dilution of 1:1,000 overnight at 4°C.

Techniques: Expressing