Journal: iScience

Article Title: Histone malonylation is regulated by SIRT5 and KAT2A

doi: 10.1016/j.isci.2023.106193

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-phospho-ACC (Ser79) , Cell Signaling Technology , Cat. #3661, RRID: AB_330337.

Techniques: Recombinant, Plasmid Preparation, Software

Journal: iScience

Article Title: Histone malonylation is regulated by SIRT5 and KAT2A

doi: 10.1016/j.isci.2023.106193

Figure Lengend Snippet:

Article Snippet: Before SDS-PAGE, protein samples were heated at 70°C for 5 min. Antibodies used are listed as follows: anti-β-tubulin (Cell Signaling Technology, Cat. #2128), anti-SIRT5 (Cell Signaling Technology, Cat. #8782), anti-acetyl-lysine (Cell Signaling Technology, Cat. #9814), anti-malonyl-lysine (PTM Biolabs, Cat. #PTM-901), anti-succinyl-lysine (PTM Biolabs, Cat. # PTM-401), anti-glutaryl-lysine (PTM Biolabs, Cat. #PTM-1151), anti-ACC (Cell Signaling Technology, Cat. #3676), anti-phospho-ACC (Ser79) (Cell Signaling Technology, Cat. #3661), anti-H2B (Cell Signaling Technology, Cat. #12364), anti-KAT2A (Cell Signaling Technology, Cat. #3305), anti-HSP60 (Abcam, Cat. #ab59457), anti-Lamin A/C (Cell Signaling Technology, Cat. #2032), anti-COX IV (Abcam, Cat. #ab16056), and anti-GAPDH (Cell Signaling Technology, Cat. #5174).

Techniques: Recombinant, Plasmid Preparation, Software

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Targeted Delivery of Stk25 Antisense Oligonucleotides to Hepatocytes Protects Mice Against Nonalcoholic Fatty Liver Disease

doi: 10.1016/j.jcmgh.2018.12.004

Figure Lengend Snippet: Downstream targets and upstream activators of STK25. ( A ) Liver protein lysates from high-fat–fed mice treated with GalNAc- Stk25 ASO vs GalNAc-control ASO (12.5 mg/kg/wk) were analyzed by Western blot using antibodies specific for total ACC or phospho-ACC (Ser79), with pan-actin used as a loading control. ( B and C ) Liver lysates were collected from male C57BL/6J mice ( B ) fed a normal chow diet and fasted for 12 hours vs re-fed or ( C ) fed a normal chow diet vs a high-fat diet for 18 weeks. The lysates were immunoprecipitated with anti-STK25 antibody and analyzed by Western blot using antibodies specific for total STK25 or phospho-threonine. Immunoprecipitation of liver lysates from Stk25 knockout mice was included as a negative control. Protein levels were analyzed by densitometry and data are shown as the total and phospho-protein abundance, as well as the ratio of phospho-protein to total protein. Representative Western blots are shown. Data are means ± SD from 6–8 mice per group. Cntr, control. * P < .05.

Article Snippet: The detection of specific proteins used the following primary antibodies: anti-STK25, anti-PCNA, anti-Ki67, anti-ACC (#3662; Cell Signaling Technology, Boston, MA), anti–phospho-ACC (Ser79; #3661; Cell Signaling Technology), anti–phospho-threonine (13-9200; Invitrogen), anti–pan-actin (sc-8432; Santa Cruz Biotechnology), anti–glyceraldehyde-3-phosphate-dehydrogenase (MA5-15738; Invitrogen), anti–β-actin (ab8226; Abcam), and horseradish-peroxidase–conjugated secondary antibodies anti-rabbit IgG (#7074; Cell Signaling Technology), anti-mouse IgG (#7076; Cell Signaling Technology), anti-rat IgG (#7077; Cell signaling Technology), and VeriBlot for IP Detection Reagent (ab131366; Abcam).

Techniques: Western Blot, Immunoprecipitation, Knock-Out, Negative Control

Journal: BioMed Research International

Article Title: Green Tea Polyphenol Epigallocatechin-3-Gallate Enhance Glycogen Synthesis and Inhibit Lipogenesis in Hepatocytes

doi: 10.1155/2013/920128

Figure Lengend Snippet: Effects of GTP-EGCG on expressions of phospho-AMPK α (Thr172) and phospho-ACC (Ser79) in HepG2 cells. Cell lysates were separated on 7.5% SDS-PAGE and incubated with 1 : 1000 phospho-AMPK α (Thr172), AMPK α , phospho-ACC (Ser79), and ACC (Cell Signaling Technology Inc, USA) β -actin was used as a loading control. A representative western blot analysis from 5 independent experiments is shown (a). Data ((b) and (c)) are expressed as mean ± S.E. ( n = 5), * P < 0.05, compared with HepG2 cells with 30 mM glucose.

Article Snippet: HepG2 lysates were subjected to 7.5% SDS-polyacrylamide gel electrophoresis then transferred to 0.45 μ M polyvinyldene difluoride (PVDF) membrane and immunoblotted with primary antibodies phospho-GSK3 β (Ser9), GSK3 β , phospho-GS (Ser641), GS, phospho-AMPK α (Thr172), AMPK α , phospho-ACC (Ser79), ACC (Cell Signaling Technology Inc, MA, USA), and β -actin (Santa Cruz, CA, USA) at 1 : 1000 dilution and secondary antibodies (Santa Cruz, CA, USA) at 1 : 10000 dilution.

Techniques: SDS Page, Incubation, Western Blot

Journal: PLoS ONE

Article Title: Regulation of FOXOs and p53 by SIRT1 Modulators under Oxidative Stress

doi: 10.1371/journal.pone.0073875

Figure Lengend Snippet: (A) Representative immunoblots of phospho-AMPK (Thr172), total AMPK, phospho-acetyl-CoA carboxylase (ACC) (Ser79), total ACC, and GAPDH in C2C12 cells treated with vehicle or 60 µM antimycin A (AA) with or without 30 µM resveratrol (RSV). (B) Quantification of phospho-AMPK (Thr172) level normalized to GAPDH (N = 5). (C) Quantification of phospho-ACC (Ser79) level normalized to GAPDH (N = 5). n.s. = not significant.

Article Snippet: The antibodies used were anti-active caspase 3 rabbit polyclonal (Abcam, Cambridge, MA, USA), anti-SOD2 rabbit polyclonal (Millipore), anti-acetyl-p53 rabbit polyclonal (detecting acetyl-Lys 379 on mouse p53 and acetyl-Lys 382 on human p53), anti-p53 mouse monoclonal, anti-FOXO1 rabbit monoclonal, anti-FOXO3a rabbit monoclonal, anti-phospho-AMPKα (Thr172) rabbit polyclonal, anti-AMPKα rabbit polyclonal, anti-phospho-acetyl-CoA carboxylase (ACC) (Ser79) rabbit polyclonal, anti-ACC rabbit polyclonal (Cell Signaling Technology, Tokyo, Japan), anti-acetyl-FOXO1 rabbit polyclonal, anti-FOXO4 goat polyclonal, anti-Bax mouse monoclonal (Santa Cruz), anti-GAPDH mouse monoclonal (Sigma Aldrich), and anti-SIRT1 rabbit polyclonal antibodies.

Techniques: Western Blot