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.