Additional file 1 of Differential effects of PGAM5 knockout on high fat high fructose diet and methionine choline-deficient diet induced non-alcoholic steatohepatitis (NASH) in mice
Creators: name: Li, Li
name: Guo, Chengcheng
name: Yu, Yue
name: Tie, Lu
name: Lu, Guotao
name: Liu, Feng
name: Han, Xueyao
name: Ji, Linong
name: Zou, Xiantong
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Method S1. Table S1. Sources of antibodies used in Western blot. Table S2. Sequences of genes detected in real-time PCR. Table S3. Sources of ELISA kit used in cytokines assay. Figure S1. Knockout of PGAM5 increased mRNA expressions of pro-inflammatory genes and decreased protein levels of antioxidative genes in BMDMs. a-d: Relative mRNA expressions of pro-inflammatory cytokines including IL-1 (a) , IL-6 (b), TNF-α (c) and IL-10 (d) measured by real-time PCR  in BMDMs. e-j: Relative protein levels of CAT (f), HO-1 (g), NRF2 (h), GPX6 (i) and SOD1 (j) measured by Western blot. *p&lt;0.05, **p&lt;0.01, ***p&lt;0.001, ****p&lt;0.0001, statistical differences between WT and KO group. #p&lt;0.05, ##p&lt;0.01, ###p&lt;0.001, ####p&lt;0.0001, statistical differences between treatments, genotypes or their interactions. ns, no significance; LPS, lipopolysaccharide; GPX6, glutathione peroxidase 6; HO-1, heme oxygenase-1; CAT, catalase; SOD1, superoxide dismutase 1; NRF2, nuclear factor erythroid 2- related factors; WT, wild type; KO, knockout. Figure S2. Knockdown of PGAM5 increased expressions of pro-fibrotic and decreased mRNA expressions of antioxidative genes in LX2 cells. After treated with TGF for 24h, a-g: Relative mRNA expressions of pro-fibrotic genes including Acta2 (a) , Col1a1 (b), Col3a1 (c), Timp1 (d) Mmp9 (e) and Mmp13 (f), and mRNA expressions of PGAM5 (g) were shown. h-k: Relative mRNA expressions of antioxidative genes including Ho-1 (h) , Nrf2 (i), Sod1 (j) and Cat (k) were shown. l-r: Relative protein levels of COL3A1 (m), MMP13 (n), α-SMA (o), MMP9 (p), COL1A1 (q) and PGAM5 (r) were detected in LX2 cells. *p &lt; 0.05, **p &lt; 0.01, ***p &lt; 0.001, ****p &lt; 0.0001, statistical differences between NC and PGAM5-/- group. #p&lt;0.05, ##p&lt;0.01, ###p&lt;0.001, ####p&lt;0.0001, statistical differences between treatments, genotypes or their interactions. COL, collagen; Mmp, matrix metalloproteases; Timp1, tissue inhibitors of Mmps; Ho-1, heme oxygenase-1; Nrf2, nuclear factor erythroid 2- related factors; Sod1, superoxide dismutase 1; Cat, catalase; NC, negative control; PGAM5-/-, PGAM5-knockdown. Figure S3. Knockdown of PGAM5 reduced TG accumulation and up-regulated antioxidative genes in 3T3L1 cells. a: TG levels measured in adipocytes. b-g: Relative protein levels of p-mTOR/t-mTOR (b-c), proteins of insulin transduction pathway including p-InsR/t-InsR and p-AKT/t-AKT (d-e) and antioxidative proteins including NRF2, CAT, HO-1 and GPX6 (f-g) were measured in 3T3L1 cells. *p &lt; 0.05, statistical differences between NC and siPGAM5 group. mTOR1, mammalian target of rapamycin complex 1; InsR, insulin receptor β; AKT, protein kinase B; NRF2, nuclear factor erythroid 2- related factors; CAT, catalase; HO-1, heme oxygenase-1; GPX6, glutathione peroxidase 6; NC, negative control; siPGAM5, PGAM5 knockdown. Figure S4. Original blots for Figs. 1–4. Figure S5. Original blots for Figs. 5, 6.