Supporting Information. Supporting Tables. S-Table 1 Primer pairs for RT-PCR. Product size. Gene Primer pairs

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Supporting Information Supporting Tables S-Table 1 Primer pairs for RT-PCR. Gene Primer pairs Product size (bp) FAS F: 5 TCTTGGAAGCGATGGGTA 3 429 R: 5 GGGATGTATCATTCTTGGAC 3 SREBP-1c F: 5 CGCTACCGTTCCTCTATCA 3 169 R: 5 CACTTCGCAGGGTCAGG 3 ACCα F: 5 AAGGGCTACCTCTAATG 3 653 R: 5 CACAAACCAGCGTCTCA 3 PPARα F: 5 AGGCTATCCCAGGCTTTGC 3 484 R: 5 CGTCTGACTCGGTCTTCTTG 3 ACO1 F: 5 TCAAGCAAAGCGAACCAG 3 421 R: 5 ACCATACCACCCACCAAC 3 MACD F: 5 CTGCGTGACAGAACCCTC 3 517 R: 5 CAACTTTCATCGCCATTT 3 VLACD F: 5 AGGCTTTGGCGGCTTTCTG 3 434 R: 5 CTCTGCTGGCACCTTGACT 3 UCP-2 F: 5 TGGGCACCATCCTAACC 3 375 R: 5 TGGCATTTCGGGCAACA 3 TNFα F: 5 ACATCTCCCTCCGGAAAGGA 3 150 R: 5 CGCCACGAGCAGGAATGAGA 3 IL-6 F: 5 GGATACCACCCACAACAG 3 518 R: 5 TGCCGAGTAGACCTCATAG 3 ERα F: 5 GGTCCAATTCTGACAATCGAC 3 318 R: 5 TTTCGTATCCCGCCTTTCATC 3 ERβ F: 5 GCACCTTGAGTCCAGAG 3 249 R: 5 TCAGTCCCACCATTAGC 3 AR F: 5 GCTCTGGCAGCAGTGAAGCA 3 252 R: 5 AGTCCCCATAGCGGCATTGC 3 CPT-1 F: 5 CTCAGCCTCTACGGCAAATC 3 377 R: 5 CTTCTTGATCAGGCCTTTGC 3 PPARγ F: 5 ATTCTGGCCCACCAACTTCGG 3 339 R: 5 TGGAAGCCTGATGCTTTATCCCCA 3 LXRα F: 5 TACAACCCTGGGAGTGAGA 3 310 R: 5 TTAGCATCCGTGGGAACAT 3 DGAT1 F: 5 GGATGGTCCCTACTATCCAG 3 210 R: 5 CCACCAGTCCCTGTAGAACT 3 DGAT2 F: 5 GGAGGCCACCGAAGTTAGCA 3 195 R: 5 CACCTCCCACCACGATGACAA 3 1

S-Table 2 Antibodies and dilutions Antibody Mol. Wt. Dilution Cat. nr. Source Company ERα 66KD 1:100 sc-542 rabbit polyclonal IgG Santa Cruz TNFα 17KD 1:200 sc-1350 goat polyclonal IgG Santa Cruz AR 87KD 1:200 sc-816 rabbit polyclonal IgG Santa Cruz UCP2 32KD 1:200 sc-6526 goat polyclonal IgG Santa Cruz FAS 207KD 1:200 sc-20140 rabbit polyclonal IgG Santa Cruz CPT1 86KD 1:200 sc-20514 goat polyclonal IgG Santa Cruz HMGCR 43KD 1:200 sc-27578 goat polyclonal IgG Santa Cruz β-actin 42KD 1:500 sc-47778 mouse monoclonal IgG Santa Cruz IL-6 26KD 1:1000 ab-6672 rabbit polyclonal IgG Abcam P-ACC 280KD 1:1000 #3661 rabbit polyclonal IgG Cell Signaling ACC 280KD 1:1000 #3676 rabbit monoclonal IgG Cell Signaling P-HMGCR 98KD 1:1000 #09-356 rabbit polyclonal IgG Millipore S-Table 3 Total cholesterol and cholesterol ester formation from the rat hepatocytes in vitro with different treatments (concentration expressed in nm/well). Medium with Palmitic acid : Oleic acid (1:2) Vehicle E2 E2+ICI DHT DHT+Flu E2+DHT Total cholesterol 46.67±6.67 13.31±3.32* 40±10.01 16.25±4.22* 42.52±8.83 21.74±7.38* Cholesterol 16.33±7.96 4.62±3.22* 18.46±5.66 7.86±5.34 20.18±9.32 9.64±4.81 ester * p< 0.05 treatments (E2, DHT and/or E2+DHT) vs. vehicle 2

Supplemental Figures S-Fig. 1 Accumulation of lipids in hepatocytes by ORO staining (A) Liver from ORX normal diet control group. (B) Liver cells with marked lipid accumulation in all 1, 2, and 3 zones in ORX HFD control group. (C) Liver from E2-treated group; zones 2 and 3 of liver lobules contain normal appearing liver cells rimmed with marked lipid changes in zone 1. (D) Liver from DHT-treated group; zone 2 and 1 of liver lobules contain normal appearing liver cells rimmed with marked lipid changes in zone 3. (E) Liver from E2+DHT-treated group; zone 2 and 3 of liver lobules contain normal appearing liver cells rimmed with also less lipid changes in zone 1. (F) Quantification of ORO staining shown as Mean±SEM, ***, P<0.001 versus HFD-C, high fat diet control. 3

S-Fig. 2 Histopathology of the liver tissues by hematoxylin and eosin (H/E) staining from WT gonad intact (non-orx) SD male rats (n=6/group) (A) Liver from the WT normal diet (ND) control group. Zonation is identical as in Fig. 1 (B) Liver cells with lipid accumulation in the HFD control. The inflammation and fatty liver cell presence is significantly less abundant than in HFD ORX liver (Fig. 1); HFD liver from the E2-treated group (C); zones 2 and 3 of liver lobules contain normal appearing liver cells rimmed with fatty changes in zone 3. HFD liver from the DHT-treated group (D) ; zones 1 and 2 of liver lobules contain normal appearing liver cells rimmed with fatty changes in zone 1. (E1-2) HFD liver from the E2+DHT-treated group, normalized liver cells in zones 1, 2 and 3 of liver lobules and scattered with some lipid accumulated liver cells. 4

S-Fig. 3 Genes for triglyceride synthesis in SD rat liver tissues No significant changes could be observed in any of the genes involved in TG synthesis among the treatment groups. None of these above shown treated groups versus HFD-C (high fat diet control) were statistically significant. S-Fig. 4 Relative densitometry analysis of the alterations of phosphorylation related to protein levels for P-ACC/total ACC and P-HMGCR/total HMGCR levels after different treatments (calculated from Fig. 3A). The densitometric values were pooled from three animals in each group and repeated three times by Western blot, and are shown as Mean±SEM. *, p<0.05, ***, p<0.001 versus HFD-C, high fat diet control. 5

5A) 5B) S-Fig. 5 Expression levels for genes involves in Fatty acid synthesis (A) and β-oxidation (B) in hepatocytes with different treatments. None of these above shown treated groups versus HFD-C (high fat diet control) were statistically significant. 6

S-Fig. 6 Human data for serum lipid profile parameters from milder/subnormal (regarded as control) (n=5) and severe (n=5) steatotic liver samples. None of the above values are statistically significant. 7

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