Figure 3
Influence of identified proteins and ATP on MSC proliferation. (A-C) MSC were seeded in 5% pHPL, tPRP and FBS, respectively, and stimulated for 3 days with: (A) 125, 250 and 500 μg/ml fibrinogen, (B) 20, 40 and 80 μg/ml apoA1 and (C) 45, 90, 180, and 106 nM ATP. Cell counts were measured using the CellTiter-Glo assay and then normalized to the unstimulated control to calculate relative cell count values. Symbols indicate statistically significant diffences between: * stimulation; + supplements; # MSC sources; (one symbol p < 0.05; two symbols p < 0.01). (D) Western blot verification of differentially expressed proteins, fibrinogen and apo-A1 in pHPL and tPRP (2.5 μg protein/lane, eight different batches, respectively). Functional platelet interaction network analysis for fibinogen gamma (E) and apolipoprotein A1 (F) was performed using PlateletWeb and subnet extraction according to Boyanova et al. [20] (white circles denote platelet proteins with no detected phosphorylation sites; black circles denote platelet proteins with phosphorylation sites detected in platelets; grey circles denote platelet proteins with phosphorylation sites detected in human cells).