Fig. 6: Loss of Mlkl influences platelet activation and results in impaired haemostasis.

a Tail bleeding time into 37 °C saline (3-mm tail amputation) in WT (n = 17) and Mlkl^−/− (n = 15) mice. Maximal time was set as 600 s. The bleeding time was determined as the time from the tail amputation to the moment the blood flow stopped for more than 2 min. Following cessation of bleeding for 2 min, the incision was monitored for a further 2 min, and bleeding within this period classified as re-bleeding. The length of re-bleeding time is not included in the two first groups, but shown separately as “re-bleeding”. Each symbol represents an individual mouse. Data is combined from two independent experiments. Male and female mice were included. Data are presented as median. b Blood loss assessed by haemoglobin levels in WT and Mlkl^−/− mice; WT n = 17 and Mlkl^−/− n = 15. Agonist-induced platelet activation in WT and Mlkl^−/− platelets determined by c integrin activation (JON/A) or d P-selectin positive platelets by flow cytometry, statistically-significant differences are shown relative to WT control platelets at the indicated concentration of agonist. Platelets were washed and counts adjusted before 20 min incubations with ADP (37 °C), Convulxin (room temperature (RT)), PAR4-AP (RT) and Thrombin (RT) at the indicated concentrations, n = 3–5 mice per genotype. Data are presented as mean ± SEM, student’s unpaired t-test, *P < 0.05; **P < 0.005; ***P < 0.001, ns; not significant.