Cd13 bv421

Integrin expression in response to APAP. To assess correlation of z-alpha with modulation of expression of integrins, adhesion molecules and other markers in response to APAP hepatotoxicity, remaining adherent HepaRG cells were recovered using TrypLE™ cell-dissociation reagent (Life Technologies), and stained using combinations of directly conjugated monoclonal antibodies: CD29-BV510 (563513), CD49f-BV421 (5625820, CD49d-FITC (580840), CD49c-PE, CD166-PE (559263) (all BD Biosciences, Oxford, UK); CD49a-APC-Vio770 (130-101-406), CD44-APC-Vio770 (130-099-149), CD49b-PE-Vio770 (130-100-328), CD90-PE-Vio770 (130-099-295), CD49e-APC (130-097-221), (all Miltenyi Biotec, Surrey, UK); CD13-BV421 (301716; Biolegend), CD54-FITC (mhcd5401; Caltag, Buckingham, UK). Cells were incubated with optimal concentration of antibodies (1/50) at 4 °C for 20 minutes, washed twice and resuspended in PBS containing 0.1% BSA and 0.1% sodium azide. Unstained cells were included as controls, and dead cells and debris were excluded from the analysis, based on scatter characteristics. Data for at least 10,000 live events per sample were acquired using a MACSQuant Analyzer (Miltenyi Biotec) and analyzed using FlowJo version 9.6.7 software (Flowjo LLC). Data is expressed as relative mean fluorescence intensity (MFI), and % positive staining.