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. Author manuscript; available in PMC: 2021 Sep 1.
Published in final edited form as: J Hepatol. 2020 Mar 24;73(3):616–627. doi: 10.1016/j.jhep.2020.03.023

Fig. 2. MLKL expression, phosphorylation and oligomerization in livers of FFC diet fed mice.

Fig. 2.

Expression of Mlkl mRNA in livers from (A) Rip3+/+ and Rip3−/− and (B) Mlkl+/+ and Mlkl−/− mice was assessed by qRT-PCR and normalized to 18S rRNA. MLKL and RIP3 protein in liver lysates from (C) Rip3+/+ and Rip3−/− and (D) Mlkl+/+ and Mlkl−/− mice was assessed by western blot and normalized to b-actin. Liver lysates were isolated from Rip3−/− mice on chow or FFC diet as a negative control for the RIP3 antibody. (E) Paraffin-embedded livers were de-paraffinized followed by phospho-MLKL staining. Images were acquired using a 10× objective. Representative images are shown (C–E). Values represent means ± SEM. Values with different superscripts are significantly different from each other, n = 3–6 per group, p <0.05, assessed by ANOVA. (F) Plasma membranes and (G) subcellular fractions (cytosol, P10: 10,000 g pellet) were isolated from liver tissues, resolved by non-reducing PAGE and probed with antibody to MLKL. A longer exposure of the PM fraction is shown in panel G. Images are representative on n = 3 isolations. FFC, high-fat, high-fructose, high-cholesterol; PM, plasma membrane; qRT-PCR, quantitative reverse transcription PCR.