Integrin α4β1+ /α9β1+ HEK293 cell line generation
HEK293A cells were transfected with 2 µg integrin-α4 plasmid using lipofectamine 2000 (Thermofisher) and selected using 800 µg mL-1 geneticin 48 hours post transfection. Cells were diluted to single cell to isolate individual colonies and clones expressing integrin-α4 were identified using an integrin-α4 antibody (Santa Cruz cat# sc-365209). HEK293A cells were transfected using the NEPA21 Electroporator system (Nepagene). Cells were transfected with 10 µg integrin α9-GFP plasmid in OptimMEM®. After 48 hours, cells were selected using 500 µg mL-1 geneticin selection. Cells were diluted to single cell to isolate individual clones, with clones expressing integrin α9-GFP identified by fluorescent microscopy.