At 24h post-infection, we found that the total protein against all three necroptosis biomarkers (MLKL, RIP3 and RIP1) and their phosphorylated forms (p-MLKL, p-RIP1 and p-RIP3) were increased in L. interrogans infected splenocyte lysates (Figures 5A–G), whereas only MLKL and p-MLKL was increased in L. biflexa infected lysates (Figures 5B, E). This evidence concerns the gene MLKL and infection.