Intensity scatter plots comparing FH with VWF do not show colocalization or a single correlation.

<p>HUVECs were treated and stained for FH (red) and VWF (green) as detailed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121994#pone.0121994.g001" target="_blank">Fig. 1AB</a>. Images are shown without the DAPI-stained nuclei. (A) FH detection with mouse anti-FH + goat anti-mouse AF-647 (red); (B) VWF detection with rabbit anti-VWF + chicken anti-rabbit AF-488 (green); and (C) Merged image of (A) and (B) detecting FH and VWF. The correlation coefficients, Pearson’s (PCC) and Manders’ (M1 and M2), for these 2 images are on the left bottom of the image. (D) The intensity scatter plot of the merged image in (C) showing 2 linear independent relationships of the data. The scattering of points along each of the axes indicates a low degree of signal overlap. The intensity delta is the distance from either outer color line to the plot’s center of density line and defines the area considered for colocalization. An intensity delta of 30 pixels was used for all of the colocalization measurements. The inset in (C) shows an enlargement of the plot’s origin.