integrin α6β4  (Sino Biological)

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