Fig. 7: Transient protection from LPS-induced thrombocytopenia in Caspase-8Pf4Δ/Pf4Δ mice. | Cell Death & Disease

Fig. 7: Transient protection from LPS-induced thrombocytopenia in Caspase-8Pf4Δ/Pf4Δ mice.

From: The necroptotic cell death pathway operates in megakaryocytes, but not in platelet synthesis

Fig. 7

a Western blot analysis of protein lysates generated from floxed control, Caspase-8Pf4Δ/Pf4Δ, MlklPf4Δ/Pf4Δ and Caspase-8Pf4Δ/Pf4Δ MlklPf4Δ/Pf4Δ mice. Purified blood platelets and BSA gradient purified cultured BM megakaryocytes probed with antibodies to Caspase-8 and MLKL. GAPDH was used as a loading control. b Megakaryocyte numbers per FOV in H&E-stained sections of BM from Caspase-8Pf4Δ/Pf4ΔMlklPf4Δ/Pf4Δ (n = 5) and Caspase-8fl/flMlklfl/fl control mice (n = 3). c Representative images of H&E stained sternal sections from Caspase-8Pf4Δ/Pf4ΔMlklPf4Δ/Pf4Δ and Caspase-8fl/flMlklfl/fl control mice. Megakaryocytes are indicated by asterisks. d Platelet counts at 2 h and 6 h post I.P. administration of LPS at 5 mg/ml. Mice were analysed in cohorts and blood was collected by cardiac puncture. 2 h LPS; Floxed control (WT) n = 3, Caspase-8Pf4Δ/Pf4Δ n = 4, MlklPf4Δ/Pf4Δ n = 4 and Caspase-8Pf4Δ/Pf4Δ MlklPf4Δ/Pf4Δ n = 3. 6 h LPS; WT n = 4, Caspase-8Pf4Δ/Pf4Δ n = 4, MlklPf4Δ/Pf4Δ n = 6, and Caspase-8Pf4Δ/Pf4Δ MlklPf4Δ/Pf4Δ n = 4. Platelet counts from 1 representative experiment. The experiment has been repeated 3 times. Statistically-significant differences are indicated relative to prebleed of WT, Caspase-8Pf4Δ/Pf4Δ, MlklPf4Δ/Pf4Δ and Caspase-8Pf4Δ/Pf4Δ MlklPf4Δ/Pf4 mice or where indicated. e Megakaryocyte numbers per FOV in H&E-stained sections of BM from LPS treated wild-type, Caspase-8Pf4Δ/Pf4Δ, MlklPf4Δ/Pf4Δ and Caspase-8Pf4Δ/Pf4Δ MlklPf4Δ/Pf4Δ mice. 2 h LPS WT n = 5, Caspase-8Pf4Δ/Pf4Δ n = 5, MlklPf4Δ/Pf4Δ n = 4 and Caspase-8Pf4Δ/Pf4Δ MlklPf4Δ/Pf4Δ n = 4. 6 h LPS; WT n = 5, Caspase-8Pf4Δ/Pf4Δ n = 5, MlklPf4Δ/Pf4Δ n = 6, and Caspase-8Pf4Δ/Pf4Δ MlklPf4Δ/Pf4Δ n = 4. Data are presented as mean ± s.d, Two-way ANOVA relative to untreated for WT, Caspase-8Pf4Δ/Pf4Δ, MlklPf4Δ/Pf4Δ and Caspase-8Pf4Δ/Pf4Δ MlklPf4Δ/Pf4Δ or where indicated with the Bonferroni multiple comparison test, *P < 0.05; **P < 0.005; ***P < 0.0005, ****P < 0.0001.

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