Blocking antibody against tTG and expression of antisense tTG construct inhibit spreading of TPA-stimulated THP-1 cells on Fn.
The 5 × 103 THP-vector and THP-antisense cells, either untreated (−TPA) or treated for 1 hour with 150 ng/mL TPA (+TPA), were plated for 12 hours at 37°C in serum-free AIM-V medium containing 0.5% BSA on Fn-coated wells. Some samples of THP-vector cells were preincubated for 1 hour with function-blocking polyclonal anti-tTG antibody, blocking mAbs against β1-, β2-, or β3-integrins, antibody against FXIIIA, or control nonimmune IgG, before plating on Fn. The antibodies were present in the medium during the assay. Adherent cells were washed with PBS, fixed with paraformaldehyde, stained with Coomassie brilliant blue, and photographed. Bar indicates 50 μm.
