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Zebrafish G6fL is required for haemostasis. A) A novel α-zebrafish G6fL antibody was used to determine knock-down of G6fL in morpholino (MO) treated larvae. 48 hrs post injection, 3 dpf larvae were lyophilised and lysed. Proteins were separated by SDS-PAGE and Western blotted with the antibody. The membrane was subsequently stripped and reprobed with an α-actin antibody to confirm equal loading. B) Control and G6fL MO treated adult fish were bled, and the blood used for a plate tilt aggregation assay. Time to aggregation (TTA) was measured following addition of either ADP (20 μM) or collagen (100 μg/ml). C) Control and G6fL MO treated larvae were immobilised on a microscope stage and then injured with a laser in the vessel wall of either veins or arteries (D). Time to occlusion (TTO) of vessels was measured for up to 2 min at which time the experiment was ceased. Statistical significance was calculated with a Student′s t-test (*P>0.05).
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