Journal: Redox Biology

Article Title: Vinyl chloride-induced interaction of nonalcoholic and toxicant-associated steatohepatitis: Protection by the ALDH2 activator Alda-1

doi: 10.1016/j.redox.2019.101205

Figure Lengend Snippet: Alda-1 attenuated hepatic inflammation and oxidative stress. A: Representative photomicrographs of CAE (neutrophils, purple) and MDA (index of oxidative stress, brown) are shown at 200x magnification. B: CAE-positive cells were counted and graphed as positive cells per 100 hepatocytes. Hepatic MDA levels (nmol/mg protein) were measured using a thiobarbituric acid reactive substances (TBARS) assay, as described in Material and Methods. a , p < 0.05 compared to LFD control, b , p < 0.05 compared to absence of VC, c , p < 0.05 compared to absence of Alda-1. Samples size per group n = 8–10.

Article Snippet: Hepatic MDA levels were quantified by determination of thiobarbituric acid reactive substances (TBARS) concentration using a commercially available kit (R&D systems, Minneapolis, MN) according to the manufacturer's instructions.

Techniques: TBARS Assay, Control

Journal: Cell metabolism

Article Title: Pyruvate carboxylase mediated anaplerosis promotes antioxidant capacity by sustaining TCA cycle and redox metabolism in liver

doi: 10.1016/j.cmet.2019.03.014

Figure Lengend Snippet: Expression levels of genes whose products are involved in A) inflammation and B) oxidative stress in 12 and 18-week HFD mice normalized to cyclophilin B; n=4-6. NADPH/NADP+ in snap frozen livers of fasted mice, indicated by C) the malate/pyruvate ratio and the malic enzyme equilibrium constant, or D) total NADP+ and NADPH ratio. E) Total amount of hepatic glutathione in the reduced state; n=4-6. F) Lipid peroxidation measured by the TBARS assay; n=4-6. G) Correlation between hepatic inflammation (IL6 expression) and NADPH/ NADP+ ratio. H) Schematic illustrating the effect of LPCKO on hepatic inflammation and oxidative stress. Data expressed as mean ± SEM. * Indicates p<.05 by Student’s t-test or 2-way ANOVA. Significance of relationship determined by Spearman correlation. See also Figure S7.

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-Pcb (Pyruvate Carboxylase) Santa Cruz Biotech Cat# sc-67021 Rabbit anti-Ppib (Cyclophilin B) Abcam Cat# Ab16045 Rabbit anti-Akt Cell Signaling Cat# 9272 Rabbit anti-Phospho-Akt (Thr308) Cell Signaling Cat# 2965 Chemicals, Peptides, and Recombinant Proteins U- 13 C Propionate Cambridge Isomiddlees Cat# CLM-1865 2 H 2 O Cambridge Isomiddlees Cat# DLM-4-1000 U- 13 C Lactate Cambridge Isomiddlees Cat# CLM-1579 U- 13 C Glucose Cambridge Isomiddlees Cat# CLM-1396 U- 13 C Pyruvate Sigma-Aldrich Cat# 490717 Sodium pyruvate Sigma-Aldrich Cat# P2256 Sodium propionate Sigma-Aldrich Cat# P1880 L-(+)-Lactic acid solution Sigma-Aldrich Cat# L1875 Glycerol Sigma-Aldrich Cat# G5516 Glutamine Sigma-Aldrich Cat# G8540 Aspartic Acid Sigma-Aldrich Cat# A7219 Bovine Serum Albumin Sigma-Aldrich Cat# A3803 Palmitic Acid Sigma-Aldrich Cat# P5585 Palmitoleic Acid Sigma-Aldrich Cat# P9417 Stearic Acid Sigma-Aldrich Cat# S4751 Oleic Acid Sigma-Aldrich Cat# O1008 Linoleic Acid Sigma-Aldrich Cat# L1376 γ-Linoleic Acid Sigma-Aldrich Cat# L2378 Decosahexanoic acid Sigma-Aldrich Cat# D2534 Amberlite Resin Sigma-Aldrich Cat# A9960 Dowex Resin Sigma-Aldrich Cat# 217506 70% Perchloric Acid Sigma-Aldrich Cat# 244252 Ethyl Acetate EMD Millipore Cat# EX0245 ADP Sigma-Aldrich Cat# A2754 CCCP Sigma-Aldrich Cat# C2759 Oligomycin Sigma-Aldrich Cat# O4876 Antimycin A Sigma-Aldrich Cat# A8674 Succinate Sigma-Aldrich Cat# S2378 Malate Sigma-Aldrich Cat# 240176 Glutamate Sigma-Aldrich Cat# 49621 Amyloglucosidase Sigma-Aldrich Cat# A7420 Critical Commercial Assays Ammonia Assay Kit Abcam Cat# ab83360 Urea Assay Kit Abcam Cat# ab83362 GSH/GSSG Ratio Detection Assay Kit Abcam Cat#ab138881 Seahorse XFe96 FluxPak mini Agilent Cat# 102601–100 iScript cDNA Synthesis Kit BioRad Cat# 1708890 Mouse Glucagon Elisa Kit Crystal Chem Cat# 81518 Ultra-Sensitive Mouse Insulin ELISA Crystal Chem Cat# 90080 TBARS Assay Kit Cayman Chemical Cat# 10009055 Creatinine Assay Kit Cayman Chemical Cat# 500701 Cell Lytic MT Cell Lysis reagent Sigma Aldrich Cat# C3228 Citrate Synthase Activity Assay Sigma-Aldrich Cat# CS0720 Glucose (HK) Assay Kit Sigma-Aldrich Cat# GAHK20 Triglyceride Determination Kit Sigma-Aldrich Cat# TRO100 ALT Activity Assay Sigma-Aldrich Cat# MAK052 AST Activity Assay Sigma-Aldrich Cat# MAK055 RNA STAT-60 Tel-Test Cat# Cs-111 Pierce BCA Protein Assay Thermo Fisher Cat# 23225 SuperSignal West Pico Chemiluminescent Substrate Thermo Fisher Cat# 34080 Autokit Total Ketones Wako Diagnostics Cat# 411-73401 HR Series NEFA Reagent Wako Diagnostics Cat# 955-34693 Deposited Data NA Experimental Models: Organisms/Strains Mouse: Pyruvate carboxylase floxed Made in lab.

Techniques: Expressing, TBARS Assay

Journal: Cell metabolism

Article Title: Pyruvate carboxylase mediated anaplerosis promotes antioxidant capacity by sustaining TCA cycle and redox metabolism in liver

doi: 10.1016/j.cmet.2019.03.014

Figure Lengend Snippet: Key Resource Table

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-Pcb (Pyruvate Carboxylase) Santa Cruz Biotech Cat# sc-67021 Rabbit anti-Ppib (Cyclophilin B) Abcam Cat# Ab16045 Rabbit anti-Akt Cell Signaling Cat# 9272 Rabbit anti-Phospho-Akt (Thr308) Cell Signaling Cat# 2965 Chemicals, Peptides, and Recombinant Proteins U- 13 C Propionate Cambridge Isomiddlees Cat# CLM-1865 2 H 2 O Cambridge Isomiddlees Cat# DLM-4-1000 U- 13 C Lactate Cambridge Isomiddlees Cat# CLM-1579 U- 13 C Glucose Cambridge Isomiddlees Cat# CLM-1396 U- 13 C Pyruvate Sigma-Aldrich Cat# 490717 Sodium pyruvate Sigma-Aldrich Cat# P2256 Sodium propionate Sigma-Aldrich Cat# P1880 L-(+)-Lactic acid solution Sigma-Aldrich Cat# L1875 Glycerol Sigma-Aldrich Cat# G5516 Glutamine Sigma-Aldrich Cat# G8540 Aspartic Acid Sigma-Aldrich Cat# A7219 Bovine Serum Albumin Sigma-Aldrich Cat# A3803 Palmitic Acid Sigma-Aldrich Cat# P5585 Palmitoleic Acid Sigma-Aldrich Cat# P9417 Stearic Acid Sigma-Aldrich Cat# S4751 Oleic Acid Sigma-Aldrich Cat# O1008 Linoleic Acid Sigma-Aldrich Cat# L1376 γ-Linoleic Acid Sigma-Aldrich Cat# L2378 Decosahexanoic acid Sigma-Aldrich Cat# D2534 Amberlite Resin Sigma-Aldrich Cat# A9960 Dowex Resin Sigma-Aldrich Cat# 217506 70% Perchloric Acid Sigma-Aldrich Cat# 244252 Ethyl Acetate EMD Millipore Cat# EX0245 ADP Sigma-Aldrich Cat# A2754 CCCP Sigma-Aldrich Cat# C2759 Oligomycin Sigma-Aldrich Cat# O4876 Antimycin A Sigma-Aldrich Cat# A8674 Succinate Sigma-Aldrich Cat# S2378 Malate Sigma-Aldrich Cat# 240176 Glutamate Sigma-Aldrich Cat# 49621 Amyloglucosidase Sigma-Aldrich Cat# A7420 Critical Commercial Assays Ammonia Assay Kit Abcam Cat# ab83360 Urea Assay Kit Abcam Cat# ab83362 GSH/GSSG Ratio Detection Assay Kit Abcam Cat#ab138881 Seahorse XFe96 FluxPak mini Agilent Cat# 102601–100 iScript cDNA Synthesis Kit BioRad Cat# 1708890 Mouse Glucagon Elisa Kit Crystal Chem Cat# 81518 Ultra-Sensitive Mouse Insulin ELISA Crystal Chem Cat# 90080 TBARS Assay Kit Cayman Chemical Cat# 10009055 Creatinine Assay Kit Cayman Chemical Cat# 500701 Cell Lytic MT Cell Lysis reagent Sigma Aldrich Cat# C3228 Citrate Synthase Activity Assay Sigma-Aldrich Cat# CS0720 Glucose (HK) Assay Kit Sigma-Aldrich Cat# GAHK20 Triglyceride Determination Kit Sigma-Aldrich Cat# TRO100 ALT Activity Assay Sigma-Aldrich Cat# MAK052 AST Activity Assay Sigma-Aldrich Cat# MAK055 RNA STAT-60 Tel-Test Cat# Cs-111 Pierce BCA Protein Assay Thermo Fisher Cat# 23225 SuperSignal West Pico Chemiluminescent Substrate Thermo Fisher Cat# 34080 Autokit Total Ketones Wako Diagnostics Cat# 411-73401 HR Series NEFA Reagent Wako Diagnostics Cat# 955-34693 Deposited Data NA Experimental Models: Organisms/Strains Mouse: Pyruvate carboxylase floxed Made in lab.

Techniques: Recombinant, Detection Assay, Enzyme-linked Immunosorbent Assay, TBARS Assay, Lysis, Activity Assay, Bicinchoninic Acid Protein Assay, Software

Journal: Redox Biology

Article Title: CXCL5 inhibits excessive oxidative stress by regulating white adipocyte differentiation

doi: 10.1016/j.redox.2022.102359

Figure Lengend Snippet: Increased oxidative stress due to decreased antioxidant proteins in Cxcl5 KO mice A) Schematic diagram of proteomic analysis of WT and Cxcl5 KO mice fed an NFD or HFD. B) Heatmap with a pattern that increases in WT HFD but decreases in Cxcl5 KO HFD. and C) GO analysis of proteins that decreased in Cxcl5 KO HFD eWAT among the list of proteins increased more than 1.2 times in WT HFD eWAT compared to WT eWAT. D) Lipid peroxidation evaluated by the TBARS assay in eWAT of WT or Cxcl5 KO mice fed NFD and HFD. E) Western blot of phosphorylation AKT Ser473 and phosphorylation JNK in eWAT proteins from WT or Cxcl5 KO mice fed NFD and HFD. F) Western blot of phosphorylation AKT Ser473 and total AKT in 3T3-L1 cells treated with CXCL5 or insulin or treated together. G) mRNA expression of Il-6 , Tnfα , Adipoq , Glut4, and Glut5 . The 36b4 gene was used for internal controls for RNA and protein. Statistical analysis was performed using two-tailed unpaired Student's t-tests. *p < 0.05 and **p < 0.01.

Article Snippet: A TBARS assay was performed using a TBARS Assay Kit (BO-TBR-200, BIOMAX, KR) to measure the level of lipid peroxidation in the adipose tissue of the mice fed an HFD.

Techniques: TBARS Assay, Western Blot, Phospho-proteomics, Expressing, Two Tailed Test