Figure 6: Tick HSP-70 like protein participates in variable fibrinogenolysis.

Fibrinogenolysis assays performed with salivary gland lysates prepared from ticks fed on immunocompetent animals in the presence of 100 μM VER155008 (HSP70 inhibitor) or equal volume of mock control and assayed at indicated time points. 12% SDS-PAGE gel (Bio-Rad) was used in the assay. Fibrinogenolysis assays with VER155008 were performed at least three times. Solid arrow indicates increased Aα chain fibrinogen degradation and dotted arrow indicates increased level of fibrinogen-degraded product at 30 min time point in mock control in comparison to HSP70-inhibitor-treated samples. (B) Densitometry analysis (for image in (A) showing levels of degradation of Aα chain of fibrinogen in mock- or HSP70 inhibitor-treated samples at the indicated time points. (C) Fibrinogenolysis assays performed with salivary gland lysates prepared from ticks fed on immunocompetent animals in the presence of 50 ng of anti-HSP70 antibody or isotype control antibody and assayed at indicated time points is shown. Arrow indicates increased fibrinogen degradation in the presence of isotype-matched control antibody. 4–20% gradient SDS-PAGE gel (NuPAGE) gels were used in the analysis. Assays with Anti-HSP70 antibody was performed three times. (D) Densitometry analysis (for gel image in (C) showing levels of degradation of Aα chain of fibrinogen in isotype- or HSP70 antibody-treated samples at the indicated time points. In (B and D), the levels of Aα chain degradation for each sample was measured relative to the respective levels of Aα chain at 0 min time point.