integrin α6β4  (R&D Systems)

Journal: International Journal of Molecular Sciences

Article Title: HIV-1 Tat Protein Enters Dysfunctional Endothelial Cells via Integrins and Renders Them Permissive to Virus Replication

doi: 10.3390/ijms22010317

Figure Lengend Snippet: Blockade of Tat entry in IC-HUVEC by antibodies directed against the α5β1, αvβ3, and/or αvβ5 integrin. In ( A ), activated HUVEC were suspended and then incubated for 2 h with 5 μg/mL of monoclonal antibodies (mAb) directed against CD31 (upper left panel), α5β1 (upper right panel), or αvβ3 (lower left panel), or with anti-α5β1 and anti-αvβ3 mAb, combined together at 5 μg/mL each (lower right panel). In ( B ), IC-HUVEC were suspended and then incubated for 2 h with 5 μg/mL of anti-α6β4 mAb (upper left panel), or with 5 μg/mL of anti-αvβ5 mAb, alone (upper right panel) or combined with anti-α5β1 and anti-αvβ3 mAb (lower panel). For both ( A , B ), IC-HUVEC incubated with Ab dilution buffer (PBS-0.1% BSA) were employed as controls (Nil). After incubation, IC-HUVEC was exposed for 10 min to biologically active Tat (1–1000 ng/mL) or its buffer. Intracellular Tat content was assayed by intracellular staining and flow cytometry as described. Results are expressed as the percentage of positive cells as compared to isotype-stained samples and non-permeabilized cells. Box-plot data obtained from three to four independent experiments and analyzed by the Mann–Whitney test are shown. Dots indicate individual measures. Orange plots refer to IC-HUVEC incubated with the said mAbs; blue plots refer to control IC-HUVEC.

Article Snippet: MAbs directed against the α5β1, αvβ3, αvβ5, or α6β4 integrins were obtained from Chemicon-Merck-Millipore (Darmstadt, Germany).

Techniques: Incubation, Bioprocessing, Staining, Flow Cytometry, MANN-WHITNEY, Control

Journal: Scientific Reports

Article Title: Enteric Species F Human Adenoviruses use Laminin-Binding Integrins as Co-Receptors for Infection of Ht-29 Cells

doi: 10.1038/s41598-018-28255-7

Figure Lengend Snippet: SPR analysis of 41PB, 40PB and 5PB interactions with α6β1 and α6β4 integrins. 41PB ( A , B ), 40PB ( C , D ) and 5PB ( E , F ) were added in two-fold dilution series to immobilised integrins. Results are shown in response units. ( G ) Kinetics of the interaction between the penton bases and integrins. Each interaction was measured four times. Variance is shown as ± SD.

Article Snippet: Recombinant human integrins used for surface plasmon resonance (SPR) experiments (α6β4 and α6β1) were purchased from R&D Systems.

Techniques:

Journal: Scientific Reports

Article Title: Enteric Species F Human Adenoviruses use Laminin-Binding Integrins as Co-Receptors for Infection of Ht-29 Cells

doi: 10.1038/s41598-018-28255-7

Figure Lengend Snippet: Examples of LDV-, RGD-, collagen- and laminin-binding integrins, their ligands and their presence/location in the small intestine.

Article Snippet: Recombinant human integrins used for surface plasmon resonance (SPR) experiments (α6β4 and α6β1) were purchased from R&D Systems.

Techniques:

Journal: Breast Cancer Research : BCR

Article Title: Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function

doi: 10.1186/s13058-017-0822-9

Figure Lengend Snippet: MMP-8 expression alters α6β4 subcellular distribution and myoepithelial cell morphology. a (i) Confocal images of β6-1089 cells transfected with empty vector ( EMPTY ) wild-type matrix metalloproteinase-8 ( MMP-8 WT ) or inactive mutant MMP-8 ( EA ) and grown on Fibronectin for 24 hours then fixed and co-stained for α6β4 ( red ) and plectin ( green ). Insert: cartoon illustrating hemidesmosome ( HD ) components, indicating the integrin α6β4 associates with plectin to link the complex to the keratin cytoskeleton. (ii) Co-localisation of α6β4 and plectin was determined by quantifying the number of yellow pixels per field of view in each condition then expressed as a ratio as compared to EMPTY. β6-1089 transfected with WT demonstrated a significant increase in α6β4 and plectin co-localisation, while the β6-1089 transfected with EA exhibited no change as compared to EMPTY. b (i) Confocal images of β6-1089 cells transfected with empty vector (EMPTY) wild-type MMP-8 (WT) or inactive mutant MMP-8 (EA) and grown on Fibronectin for 24 hours then fixed and co-stained for α6β4 ( green ) and phallodin ( red ). (ii) Morphological features of the transfected β6-1089 were analysed and length of trailing fibres was noted to be altered. Analysis of the length of these fibres indicated that β6-1089 transfected with MMP-8 WT significantly reduced the fibre length as compared to β6-1089 transfected with empty vector or MMP-8 EA. (iii) Analysis of the number of these fibres per cell also demonstrated a significant reduction in β6-1089 transfected with MMP-8 WT as compared to β6-1089 transfected with empty vector or MMP-8 EA. (iv) The final morphological alteration analysed was the physical space the cells occupied, the relative area occupied by β6-1089 transfected with MMP-8 WT was significantly larger than that occupied by β6-1089 transfected with empty vector or MMP-8 EA. c (i) Confocal images of N-1089 cells transfected with siRNA targeting MMP-8 ( siMMP-8 ) or Luciferase ( siLuc ) as a control grown on Fibronectin for 24 hours then fixed and co-stained for α6β4 ( green ) and phallodin ( red ). (ii) Trailing fibre length was significantly longer in N-1089 transfected with siMMP8 as compared to siLUC. (iii) There were significantly more trailing fibres per cell in the N-1089 + siMMP8 group as compared to N-1089 transfected with siLUC. * p = 0.05 ** p = 0.01, *** p = 0.001 (Student’s t test). Error bars = SEM, n = 3 independent experiments

Article Snippet: Primary antibodies used were α6β4 integrin (Merck Millipore, Billerica, MA, USA, MAB1964) and plectin (Epitomics, Burlingame, CA, USA, 1399).

Techniques: Expressing, Transfection, Plasmid Preparation, Mutagenesis, Staining, Luciferase, Control