Organochloride pesticides impaired mitochondrial function in hepatocytes and. aggravated disorders of fatty acids metabolism

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1 Supplementary Materials Organochloride pesticides impaired mitochondrial function in hepatocytes and aggravated disorders of fatty acids metabolism Qian Liu 1,2,3*, Qihan Wang 4*, Cheng Xu 2,3, Wentao Shao 2,3, Chunlan Zhang 2,3, Hui Liu 2,3, Zhaoyan Jiang 1,4, Aihua Gu 2,3 1 Center of Gallbladder Disease, Shanghai East Hospital, Institution of Gallstone Disease, Tongji University School of Medicine, Shanghai, China, State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing, China 3 Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing, China 4 Department of Surgery, Shanghai Institute of Digestive Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China, * These authors contributed equally to this study and all should be considered as first authors. Corresponding authors: Aihua Gu: aihuagu@njmu.edu.cn Zhao-Yan Jiang: zhaoyanjiang@gmail.com

2 Supplement Table 1. Sequences of primers for RT-PCR. Supplement Table 2. Effect of p,p -DDE and β-hch on serum biochemical Supplement Table 3. Metabolomic analysis of liver tissues from mouse exposed to OCPs Supplementary Figure Legends Figure S1 Changes in body weight among treatment groups (n=8/group). Figure S2 Cell viability was analyzed after 24 h exposure to different concentrations of p, p -DDE (A) and β-hch (B). The results were expressed as a ratio relative to controls and presented as mean ± SEM of six independent experiments.

3 Supplement Table 1. Sequences of primers for RT-PCR. Gene Mouse-Acc F Mouse-Acc R Mouse-Fas F Mouse-Fas R Mouse-Scd1 F Mouse-Scd1 R Mouse-Cpt1α F Mouse-Cpt1α R Mouse-Acox1 F Mouse-Acox1 R Mouse-Scad F Mouse-Scad R Mouse-Mcad F Mouse-Mcad R Mouse-Lcad F Mouse-Lcad R Mouse-GAPDH F Mouse-GAPDH R Human-CPT1α F Human-CPT1α R Human-SCAD F Human-SCAD R Human-MCAD F Human-MCAD R Human-LCAD F Human-LCAD R Human-GAPDH F Human-GAPDH R Sequences GCCGTGGGGAAGGAAAAGT GTGGCTAGGTACTGAACAAAGAG TATCAAGGAGGCCCATTTTGC TGTTTCCACTTCTAAACCATGCT TTCTTGCGATACACTCTGGTGC CGGGATTGAATGTTCTTGTCGT ACGTTGGACGAATCGGAACA GGTGGCCATGACATACTCCC CCGTCGAGAAATCGAGAACT ATTGAGGCCAACAGGTTCCA ATGTGCCAGAGGAGCTGAGT TGATCCACTGTTGCTTCTGC AACTAAACATGGGCCAGCGA CAGCTGCGACTGTAGGTCTG GCATCAACATCGCAGAGAAA ACGCTTGCTCTTCCCAAGTA AGGTCGGTGTGAACGGATTTG TGTAGACCATGTAGTTGAGGTCA ATCAATCGGACTCTGGAAACGG TCAGGGAGTAGCGCATGGT CGGCAGTTACACACCATCTAC GCAATGGGAAACAACTCCTTCTC GGAAGCACATACCCCAGGAAT AGCTCCGTCACCAATTAAAACAT TGCAATAGCAATGACAGAGCC CGCAACTACAATCACAACATCAC TGACAACTTTGGTATCGTGGAAGG AGGCAGGGATGATGTTCTGGAGAG F: forward; R: reverse

4 Supplement Table 2. Effect of p,p -DDE and β-hch on serum biochemical Index Control p,p'-dde β-hch ALT (U/L) 38.6± ± ±5.8 AST (U/L) 78.9± ± ±8.6 TP (g/l) 56.9± ± ±0.7 ALB (g/l) 38.3± ± ±0.7 TBIL (umol/l) 1.9± ± ±1.1 ALP (U/L) 90.0± ± ±6.1 GLU (mmol/l) 11.0± ± ±0.5 BUN (mmol/l) 9.9± ± ±0.6 CREA (umol/l) 9.0± ± ±1.4 CHOL (mmol/l) 3.1± ± ±0.1 TG (mmol/l) 0.8± ± ±0.1 HDL-C (mmol/l) 2.5± ± ±0.1 LDL-C (mmol/l) 0.1± ± ±0.02 GLOB (g/l) 18.7± ± ±0.3* parameters (mean ± SEM, n = 8/group). Significance indicated by: *p < 0.05 compared with the control group.

5 Supplement Table 3. Metabolomics analysis of liver tissues from mouse exposed to OCPs Metabolites RT mass Metabolic pathway Fold changes (DDE/Ctrl) Fold changes (DDE/Ctrl) Cinnamic acid Amino acids metabolism Xanthurenic acid Tryptophan metabolism Sphinganine Sphingolipid metabolites Phenylpyruvic acid Phenylalanine metabolism PG(P-16:0/14:1(9Z)) Phospholipids metabolism PE(18:1(9Z)/0:0) Phospholipids metabolism PE(18:0/0:0) Phospholipids metabolism PC(O-18:0/0:0) Phospholipids metabolism PC(8:0/7:0)[ Phospholipids metabolism PC(14:0/0:0) Phospholipids metabolism Palmitic amide Fatty acid metabolism PA(22:1(11Z)/0:0) Phospholipids metabolism PA(20:1(11Z)/0:0) Phospholipids metabolism Homocysteine Amino acids metabolism Histidine Amino acids metabolism Uric acid Purine metabolism PI(18:1(9Z)/0:0) Phospholipids metabolism Phenylalanine Amino acids metabolism Glucose 6-phosphate Glucose metabolism Xanthurenic acid Tryptophan metabolism Xanthine Purine metabolism PS(19:0/0:0) Phospholipids metabolism PG(18:1(9Z)/0:0) Phospholipids metabolism Glucose Glucose metabolism PI(16:0/0:0) Phospholipids metabolism

6 Figure S1

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