Fig. 7.
PAI-1 disrupts actin stress fibers. MG-63 cell monolayers were plated overnight onto coverslips in complete media and then incubated for 3 hours with 20 nM of PAI-1 or PAI-1 mutants (PAI-1R and Q123K). Cells were subsequently fixed, permeabilized and stained for 1 hour with the clone 15F11 antibody for β5 integrin. Cells were then washed and labeled for an additional hour with the AlexaFluor488 derivatized anti-mouse secondary antibody and the AlexaFluor594-phalloidin. Actin and β5 integrin were visualized by indirect immunofluorescence. Scale Bar, 20 μm.