Figure 2

Bivariate dot plots of "Area Threshold 30%" representing diffuseness of staining on the X axis and "Bright Detail Intensity-FITC" representing the degree of punctuate staining on the Y axis (see Materials and Methods). MDA-MB-231 cells were exposed to anti-β4 on ice, followed by control rabbit IgG (A) or rabbit anti-mouse IgG (B) at 37°C to crosslink α6β4 for 30 min. Cells were stained with FITC-labeled anti-EGFR and nuclear stain DRAQ5 and run on the ImageStream™. Representative brightfield (BF) and fluorescent images from gated cells to the right of the diagonal in B show a diffuse cell surface distribution of EGFR (C-E), whereas representative images of gated cells to the left of the diagonal in B show a clustered distribution of EGFR (F-H).