Figure 5

Adhesion of monocytes to collagen is dependent on expression of APP. THP-1 cells were either not transfected or transfected with an individual APP siRNA duplex. A) At 24 hours post-transfection, cells were lysed with RIPA buffer, separated by 7% SDS-PAGE and Western blotted with anti-APP antibody and anti-ERK2 antibody (loading control). B) At 24 hours post-transfection, transfected cells and non-transfected cells were loaded with 5 μg/mL calcein-AM and plated on tissue culture plastic alone, poly-L-lysine, or type I collagen for 30 minutes. Fluorescence from total cell number was read prior to washing the wells. Wells were then washed and fluorescence of adhered cells was determined. Data were analyzed by unpaired ANOVA with Tukey's post-test comparison and are expressed as mean +/- SD (*** = p < 0.001 from control, ** = p < 0.001 from mock transfected cells). Values are representative of three independent experiments.