Additional Supporting Information may be found in the online version of this article.

HEP_25694_sm_SuppFig1.tif1963KSupporting Information Figure 1. GNMT deficiency promotes steatosis in young mice (A) H&E and Sudan Red staining showing mild steatosis in 3m-old GNMT−/− mice when compared to WT littermates.
HEP_25694_sm_SuppFig2.tif121KSupporting Information Figure 2. Determination of Kupffer cell activation markers in GNMT−/− mice. (A) qRT-PCR showing lower F4/80, IL-1β and IL-6 in untreated GNTM−/− mice vs. GNMT+/+ controls. **p < 0.01; (GNMT−/− vs. GNMT+/+).
HEP_25694_sm_SuppFig3.tif195KSupporting Information Figure 3. GNMT efficiency promotes NK1.1 cell depletion and NK cell cytotoxicity. (A) FACS analysis of splenic MNCs. The histogram shows NK1.1+ cells vs. CD3+. The graph represents the percentage of NK1.1+ cells vs. CD45+ cells. Cytotoxicity assay by co-culturing YAC-1 cells at the different ratios illustrated in the figure. LDH release was determined 12 hours later.
HEP_25694_sm_SuppFig4.tif99KSupporting Information Figure 4. Determination of serum levels of IFNγ and TNF after ConA (A) ELISA on serum samples confirmed higher presence of IFNγ but comparable TNF levels in GNMT−/− mice 6h after ConA when compared to WT littermates. **p < 0.01; (GNMT−/− vs. GNMT+/+).
HEP_25694_sm_SuppFig5.tif210KSupporting Information Figure 5. ConA promotes stronger NK1.1+ cell depletion in GNMT−/− vs GNMT+/+ mice FACS analysis of splenic MNCs after 3h of ConA treatment showed stronger depletion of NK1.1+CD3- and NK1.1+/CD3+ cells that expressed more AnnexinV in GNMT−/− mice compared to GNMT+/+ as shown by (A) dot plot and (B) further quantification represented in % of NK1.1+ vs CD45+ cells.
HEP_25694_sm_SuppFig6.tif4464KSupporting Information Figure 6. Adoptive transfer of TRAIL deficient MNCs cells protects the liver from GNMT deficient mice against LPS/GalN damage. ASIALO/GNMT−/− mice received TRAIL−/− or GNMT−/− splenic-MNCs. (A) Serum ALT, (B) H&E staining and TUNEL. (C) qRT-PCR (D) Western blot analysis n= 4. *P<0.05; **P<0.01 (GNMT−/−ASIALO/GNMT−/−MNCs vs. GNMT−/−ASIALO/TRAIL−/−MNCs); ##p<0.01 (GNMT−/− vs. GNMT−/− ASIALO/TRAIL−/− MNCs). Error bars represent SD.
HEP_25694_sm_SuppFig7.tif3462KSupporting Information Figure 7. Impact of TNF on hepatocyte and liver homeostasis in the absence of GNMT (A) Western blot analysis showed that 30ng/ml of TNF promotes stronger p- JNK1/2 and p-c-jun in GNMT−/−hepatocytes at 15 and 30 minutes. Densitometric quantification of western blots using p-JNK, p-c-jun (B) Comparable cleavage of PARP and expression of Bcl-xL 24 hours after TNF. Densitometric quantification of western blots using PARP antibodies. (C) Serum ALT levels 8 h after i.v. injection of TNF (D) H&E staining and TUNEL assay in liver sections confirmed the marginal impact of TNF (6μg/kg; 8h) on GNMT−/− mice (200x). (E) Western blot analysis on whole liver extracts showed stronger p-JNK1/2, p-c-jun and NOS2 expression in TNF/GNMT−/− mice. GAPDH was used as loading control. n= 4. *P<0.05; **P<0.01 (GNMT−/− vs. GNMT+/+). All data are representative of three independent experiments. Error bars represent SD.
HEP_25694_sm_SuppInfo.doc24KSupporting Information

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