Fig. 6.

Phosphoproteomic analysis of MAECs cultured at different time points on a coating of lactadherin/immobilized lactadherin.A, scheme illustrating the experimental setup: 10 μg/ml lactadherin was used to coat the plate. A clustered heatmap highlighting the (B) up- and (C) downregulated phosphosites and predicted kinases regulated by immobilized lactadherin in MAECs after 1 h. A clustered heatmap highlighting D up and E down regulated phosphosites and predicted kinases regulated by immobilized lactadherin in MAECs at 24 h. The score from 0 (dark blue) to 1 (red) shows the combined kinase-substrate score determined by the PhosR method (32) for the affected phosphosites (each row). A higher combined score (red) indicates a better match with a kinase motif (column) and the kinase-substrate phosphorylation profile of a phosphosites (row). For example, in (B), phosphosite S133 of MAP2K5 (row) can be strongly phosphorylated by GSK3B (column). A high combined score of GSK3B and MAPK3 (belonging to the same kinase group, the CMGC group (orange)) in association with a cluster of several upregulated phosphosites clearly indicates that these kinases can regulate the cellular processes induced by lactadherin at 1 h.