Protective Effects of Non-Anticoagulant Activated Protein C Variant (D36A/L38D/A39V) in a Murine Model of Ischaemic Stroke - Fig 6

<p><b>A.</b> Influence of murine wt APC and APC(36–39) alone or in combination with tPA on the oedema ratio (area of the infarcted hemisphere vs. non-infarcted contralateral hemisphere). <b>B.</b> Haemoglobin levels in the ischaemic hemisphere of mice treated with murine wt APC or APC(36–39) ±tPA. All values are expressed as mean ± SEM. * <i>P</i> < 0.05 compared to mice treated with vehicle. # <i>P</i> < 0.05 compared to mice treated with tPA.</p

Amino acid alignment of human and murine protein C.

<p>Amino acid sequence is separated into different domains and numbered according to the human sequence. Amino acid identity is highlighted in yellow. Gla residues are denoted by *. Glycan attachment sites are highlighted in green. Residues involved in the catalytic triad are highlighted in red. Regions known to be important for protein S, factor Va and PAR1 binding are boxed. The amino acids mutated in each of the protein C variants 36–39, 5A and Ca-ins are shown in red, blue and green respectively.</p

Catalytic efficiencies (k_cat/K_m) of human (h) and murine (m) APC and variant APC-mediated proteolysis of the peptide substrate, S2366.

<p>Catalytic efficiencies (k_cat/K_m) of human (h) and murine (m) APC and variant APC-mediated proteolysis of the peptide substrate, S2366.</p

Time course of inhibition of human APC variants by PCI.

<p>APC variants (20 nM) were incubated with 200 nM PCI at 37°C. At designated time points (0–20 minutes), hydrolysis of 400 μM S-2366 substrate was used to determine of the concentration of residual APC. The graph represents the percentage of residual APC activity over time and values (<i>n</i> = 3) represent the mean±SD.</p

Effect of murine wt APC or APC(36–39) with or without tPA upon functional recovery and infarct lesion at 24h post-MCAO.

<p>Male mice were subject to transient MCAO for 60 min. tPA (10mg/kg), murine wt APC or APC(36–39) (250μg/kg) either alone or in combination with tPA, or saline control, were i.v. infused 3 hours post MCAO. <b>A.</b> Neurological deficit scoring of MCAO treated animals at 24 hours. <b>B.</b> Infarct volume (mm³) in brains from animals receiving different treatments at 24 hours <b>C.</b> Representative cresyl violet staining of brain coronal sections of ischemic mice treated with vehicle (saline), tPA or tPA + APC(36–39). Infarct area is denoted by the red line. All values are expressed as mean ± SEM. * <i>P</i> < 0.05 compared to mice treated with vehicle. # <i>P</i> < 0.05 compared to mice treated with tPA.</p

Protective Effects of Non-Anticoagulant Activated Protein C Variant (D36A/L38D/A39V) in a Murine Model of Ischaemic Stroke - Fig 3

<p><b>A.</b> Representative example of thrombin generation in normal human plasma initiated with 4 pM TF measured in the presence of 2.5 nM wt APC or non-anticoagulant APC variants. <b>B-C.</b> Inhibition of TF-induced thrombin generation with increasing concentrations of either human (<b>B</b>) or murine (<b>C</b>) APC variants. The ETP in the absence of APC was taken as 100% and the ETP generated in the presence of each human (0 to 50 nM) or murine (0 to 20 nM) APC variant was expressed as a percentage of this. Values (<i>n</i> = 4) represent the mean±SD.</p