PSaV-induced necrosome formation (RIPK1/RIPK3/MLKL complex).

(A) Lysates of LLC-PK cells infected with PSaV Cowden strain (MOI = 1 TCID₅₀) or left mock-infected in the presence of 200 μM GCDCA were immunoprecipitated using an anti-pRIPK1 antibody at indicated time points. Immunoprecipitated pRIPK3 and pMLKL with pRIPK1 were determined using western blot analysis with relevant antibodies. Representative images of different gels from two independent experiments are presented. (B, C) The relative level of immunoprecipitated pRIPK3 or pMLKL with pRIPK1 was estimated by densitometric analysis. Data in panels B and C are presented as means ± standard error from two independent experiments. Differences were evaluated using the one-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001.