Fig. 5.
Collagen films were crosslinked with 100% EDC/NHS, incubated with Diaz-ES-VWFIIINle or VWFIIINle at 5 μg/ml and left in the dark or exposed to UV light for 5 min. COS-7 or HEK-293 cells, transiently transfected with DDR2-Flag, were incubated on films. For each condition, fluorescence intensity ratio of DDR2 phosphorylation over DDR2-Flag was measured. Y axis shows the ratio normalized to the value obtained with non-crosslinked collagen film. (A) Fluorescence intensity measured in COS-7 cells by confocal microscopy on 10 randomly selected fields of view for each conditions within each repeat. This graph shows pooled results from five independent data sets (n = 10 for each condition within each experiment; 1-way ANOVA, p < 0.0001). (B) Fluorescence intensity of DDR2 extracted from HEK293 cells and measured on western blot membrane. This graph shows pooled results from three independent data sets. Diaz-ES-VWFIIINle followed by UV treatment led to a significant increase in DDR2 phosphorylation, as measured by immunofluorescence or western blot (1-way ANOVA, p < 0.001).