Fig 1: MLKL level in the liver is positively correlated with human and mice liver fibrosis. (A) Quantitative RT-PCR analysis of the mRNA levels of Mlkl and fibrosis markers Vimentin, Fsp1, Desmin, Mmp2, Col1a1, and a-SMA in the liver biopsy samples of patients with fibrosis/cirrhosis (n=45). Pearson's correlation coefficient and statistical significance are listed in the plots. (B) Western blot analysis of RIP1, RIP3, MLKL, and their phosphorylated forms, together with fibrosis markers a-SMA and Vimentin in the liver biopsy samples of patients with fibrosis/cirrhosis (n=12, each lane represents one patient sample). Gapdh was used as loading control. (C) The Pearson's correlations among the protein levels of a-SMA, Vimentin, MLKL, and p-MLKL in (B). The protein levels were normalized to Gapdh in the same sample. (D) Western blot analysis of a-SMA, Vimentin, MLKL and p-MLKL in the liver of mice treated with CCl4 or vehicle (oil) three times a week for 8 weeks (n=4, each lane represents one animal sample). Gapdh was used as loading control. (E) Quantification of the blots in (D). The protein levels were normalized to Gapdh in the same sample, then normalized to vehicle control. (F) Quantitative RT-PCR analysis of Mlkl and fibrosis genes Col1a1, a-SMA, Desmin, Vimentin, Fsp1, and Pdgfrb in liver samples of mice treated with oil or CCl4 (n=4). All data are shown as Means ± SEM, *P < 0.05, **P< 0.01, ***P< 0.001 (Student's t-test).