Journal: iScience

Article Title: Exercise-acclimated microbiota improves skeletal muscle metabolism via circulating bile acid deconjugation

doi: 10.1016/j.isci.2023.106251

Figure Lengend Snippet: Exercise-acclimated microbiota treatment improves glucose tolerance in HFD-fed mice (AN) Body and tissue weights (n = 8, A–C), blood chemistry (n = 8, D and E), blood glucose concentrations during oral glucose tolerance tests (n = 8, GTT) (F) and insulin tolerance tests (ITT) (n = 8, G), glycogen content (n = 7–8, H), mRNA levels of PGC1α and COX4-1 (n = 8, I and J), AMPKα Thr172 (n = 7, K) and ACC Ser212 (n = 6–7, L) phosphorylation, membrane content of GLUT4 (n = 7, M), and COX activity (n = 8, N) in gastrocnemius muscle in recipient mice fed chow or HFD for 8 weeks. Phosphorylation levels were correlated with total content of each target in immunoblotting (K–M). The solid line shows absolute values, and the dotted line shows relative values in the ITT (G). RS; recipient from sedentary donor, RT; recipient from trained donor. ∗p < 0.05, and ∗∗p < 0.01 between groups. Results are presented as the mean ± SE.

Article Snippet: Subsequently, the blots were incubated with primary antibodies against phospho-AMPKα (Thr172), total AMPKα, phospho-ACC (Ser212), total ACC, phospho-Akt (Ser473), total Akt, phospho-Akt substrate (Ser/Thr) (AS), phospho-AS160 (Thr642), total AS160, phospho-calcium/calmodulin-dependent protein kinase II (CaMKII) (Thr286), total CaMKII, phospho-LKB1 (Ser428), total LKB1 (all from Cell Signaling Technology, Beverly, MA), glucose transporter 4 (GLUT4) (Merck Millipore, Darmstadt, Germany), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abcam, Cambridge, MA, USA).

Techniques: Activity Assay, Western Blot

Journal: iScience

Article Title: Exercise-acclimated microbiota improves skeletal muscle metabolism via circulating bile acid deconjugation

doi: 10.1016/j.isci.2023.106251

Figure Lengend Snippet:

Article Snippet: Subsequently, the blots were incubated with primary antibodies against phospho-AMPKα (Thr172), total AMPKα, phospho-ACC (Ser212), total ACC, phospho-Akt (Ser473), total Akt, phospho-Akt substrate (Ser/Thr) (AS), phospho-AS160 (Thr642), total AS160, phospho-calcium/calmodulin-dependent protein kinase II (CaMKII) (Thr286), total CaMKII, phospho-LKB1 (Ser428), total LKB1 (all from Cell Signaling Technology, Beverly, MA), glucose transporter 4 (GLUT4) (Merck Millipore, Darmstadt, Germany), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abcam, Cambridge, MA, USA).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Protein Extraction, Expressing, Software, Mass Spectrometry, Real-time Polymerase Chain Reaction

Journal: iScience

Article Title: Exercise-acclimated microbiota improves skeletal muscle metabolism via circulating bile acid deconjugation

doi: 10.1016/j.isci.2023.106251

Figure Lengend Snippet: Exercise-acclimated microbiota treatment improves glucose tolerance in HFD-fed mice (AN) Body and tissue weights (n = 8, A–C), blood chemistry (n = 8, D and E), blood glucose concentrations during oral glucose tolerance tests (n = 8, GTT) (F) and insulin tolerance tests (ITT) (n = 8, G), glycogen content (n = 7–8, H), mRNA levels of PGC1α and COX4-1 (n = 8, I and J), AMPKα Thr172 (n = 7, K) and ACC Ser212 (n = 6–7, L) phosphorylation, membrane content of GLUT4 (n = 7, M), and COX activity (n = 8, N) in gastrocnemius muscle in recipient mice fed chow or HFD for 8 weeks. Phosphorylation levels were correlated with total content of each target in immunoblotting (K–M). The solid line shows absolute values, and the dotted line shows relative values in the ITT (G). RS; recipient from sedentary donor, RT; recipient from trained donor. ∗p < 0.05, and ∗∗p < 0.01 between groups. Results are presented as the mean ± SE.

Article Snippet: Rabbit polyclonal anti-phospho ACC (Ser212) , Cell Signaling Technology , Cat#3661S; RRID: AB_330337.

Techniques: Activity Assay, Western Blot

Journal: iScience

Article Title: Exercise-acclimated microbiota improves skeletal muscle metabolism via circulating bile acid deconjugation

doi: 10.1016/j.isci.2023.106251

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-phospho ACC (Ser212) , Cell Signaling Technology , Cat#3661S; RRID: AB_330337.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Protein Extraction, Expressing, Software, Mass Spectrometry, Real-time Polymerase Chain Reaction

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: Nutrients

Article Title: L-Fucose Suppresses Lipid Accumulation via the AMPK Pathway in 3T3-L1 Adipocytes

doi: 10.3390/nu15030503

Figure Lengend Snippet: Effects of L-fucose (Fuc) on lipid metabolism-related proteins. ( A ) Time course of AMP-activated protein kinase (AMPK) phosphorylation in adipocytes treated with 20 mM Fuc. ( B , C ) Protein expression levels of p-AMPK, AMPK, p-acetyl-CoA carboxylase (ACC), and ACC in adipocytes treated with Fuc for 24 h. ( D , E ) Protein expression levels of p-hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL), and peroxisome proliferator-activated receptor (PPAR)γ1, 2 in adipocytes treated with Fuc for 24 h. Data are presented as mean ± SD ( n = 3), and statistical significance was calculated using Dunnett’s test (the group treated without Fuc vs. the groups treated with Fuc, * p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: The following antibodies were used in this study: AMPKα Rabbit (#2532S), Phospho-AMPKα Rabbit mAb (#2535S), ACC Rabbit Ab (#3662), Phospho-ACC Rabbit Ab (#3661), ATGL Rabbit Ab (#2138S), Phospho-HSL (#4126), PPARγ Rabbit mAb (#2443S), Akt Rabbit mAb (#4691S), and Phospho-Akt Rabbit mAb (#13038S); all of these were purchased from Cell Signaling Technology Japan (Tokyo, Japan). β-Actin mouse rabbit mAb (#010-27841) was purchased from Fujifilm Wako.

Techniques: Expressing

Journal: Nutrients

Article Title: L-Fucose Suppresses Lipid Accumulation via the AMPK Pathway in 3T3-L1 Adipocytes

doi: 10.3390/nu15030503

Figure Lengend Snippet: Effects of L-fucose (Fuc) and AMP-activated protein kinase (AMPK) inhibitor Compound C (CC) on lipid metabolism-related proteins. ( A , B ) Protein expression levels of p-AMPK, AMPK, p-acetyl-CoA carboxylase (ACC), and ACC in adipocytes cotreated with 10 µM CC and Fuc for 24 h. ( C , D ) Protein expression levels of p-hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL), and peroxisome proliferator-activated receptor (PPAR)γ1, 2 in adipocytes cotreated with 10 µM CC and Fuc for 24 h. Data are presented as mean ± SD ( n = 3), and statistical significance was calculated using Welch’s t -test (control vs. Fuc, CC vs. Fuc + CC, ** p < 0.01; *** p < 0.001).

Article Snippet: The following antibodies were used in this study: AMPKα Rabbit (#2532S), Phospho-AMPKα Rabbit mAb (#2535S), ACC Rabbit Ab (#3662), Phospho-ACC Rabbit Ab (#3661), ATGL Rabbit Ab (#2138S), Phospho-HSL (#4126), PPARγ Rabbit mAb (#2443S), Akt Rabbit mAb (#4691S), and Phospho-Akt Rabbit mAb (#13038S); all of these were purchased from Cell Signaling Technology Japan (Tokyo, Japan). β-Actin mouse rabbit mAb (#010-27841) was purchased from Fujifilm Wako.

Techniques: Expressing

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