Journal: Nature Communications

Article Title: NAD + repletion with niacin counteracts cancer cachexia

doi: 10.1038/s41467-023-37595-6

Figure Lengend Snippet: a Study design of the C26-F model and NA treatment with three experimental groups: control, C26-F and C26-F + NA. b Levels of NAD + represented as pmol per mg of muscle (control n = 7, C26-F n = 7, C26-F + NA n = 6). c , d Gastrocnemius (GSN) and tibialis anterior (TA) muscle wet weight normalized to initial body weight (IBW) ( n = 7 per group). e Grasping strength at the start of NA treatment and the day after every Folfox administration ( n = 7 per group). f–i Representative western blotting bands ( f ) and densitometry analysis of puromycin incorporation (SUnSET analysis) ( g ; control n = 6, C26-F n = 7, C26-F + NA n = 7), LC3B-II normalized to total LC3B ( h ; n = 7 per group) and p-AMPK Thr172 normalized to total AMPK ( i ; n = 7 per group) protein levels. GAPDH was used as loading control. j Study design of the VCM model and NA treatment with three experimental groups: control, VCM and VCM + NA. k Levels of NAD + represented as pmol per mg of muscle. l–n GSN, TA and quadriceps femoris (QUAD) muscle wet weight. o , p Representative western blotting bands ( o ) and densitometry analysis of LC3B-II normalized to total LC3B ( p ; control n = 8, VCM n = 8, VCM + NA n = 6) protein levels. GAPDH was used as loading control. Data are shown in ( b–d , g–i , k–n and p ) as means with individual values and in ( e ) as means ± SEM. Statistical analysis was performed using one-way ANOVA + Fisher’s LSD test. Original raw data are provided as a . NA niacin, A.U. arbitrary units.

Article Snippet: Membranes were blocked with 5% nonfat dry milk in Tris-buffered saline containing 0.05% Tween (TBS-Tween) and then incubated overnight with antibodies directed against specific proteins: AMPK (1:1000, 07-181, Millipore, polyclonal), p-AMPK Thr172 (1:1000, #2535, Cell Signaling, clone 40H9), GAPDH (1:10000, G8795, clone GAPDH-71.1), LC3B (1:1000, L7543, polyclonal), NRK2 (1:1000, produced in Dr. Gareth G Lavery’s lab, polyclonal), OXPHOS Antibody Cocktail (1:1000, ab110413, Abcam), PGC-1α (1:1000, AB3242, Merck Millipore, polyclonal), PINK1 (1:500, SAB2500794, polyclonal), Puromycin (1:1000, EQ0001, Kerafast, clone 3RH11), TOMM20 (1:1000, ab186735, Abcam, clone EPR15581-54) and Vinculin (1:2000, sc73614, Santa Cruz Biotechnology, clone 7F9).

Techniques: Western Blot

Journal: Diabetology & Metabolic Syndrome

Article Title: Dynamic evolution and mechanism of myocardial glucose metabolism in different functional phenotypes of diabetic cardiomyopathy — a study based on 18 F-FDG microPET myocardial metabolic imaging

doi: 10.1186/s13098-023-01038-5

Figure Lengend Snippet: The expression of p-AMPK in myocardium was decreased in diabetic mice. ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: The primary antibodies used in this study included anti-glucose transporter-1 (GLUT-1) antibody (Abcam/ab652), anti-GLUT-4 antibody (Abcam/ab33780), anti-AMP-activated protein kinase (AMPK) antibody (Abcam/ab3760), anti-phosphorylated AMPK (p-AMPK) antibody (CST/2535) and anti-GAPDH antibody (Abcam/ab8245).

Techniques: Expressing

Journal: Diabetology & Metabolic Syndrome

Article Title: Dynamic evolution and mechanism of myocardial glucose metabolism in different functional phenotypes of diabetic cardiomyopathy — a study based on 18 F-FDG microPET myocardial metabolic imaging

doi: 10.1186/s13098-023-01038-5

Figure Lengend Snippet: A schematic diagram of the molecular mechanisms involved in this study leading to decreased myocardial glucose metabolism. During the development of DCM, myocardial AMPK phosphorylation is inhibited, and its activity is reduced, accompanied by reduced GLUT-4 membrane translocation and myocardial glucose metabolism

Article Snippet: The primary antibodies used in this study included anti-glucose transporter-1 (GLUT-1) antibody (Abcam/ab652), anti-GLUT-4 antibody (Abcam/ab33780), anti-AMP-activated protein kinase (AMPK) antibody (Abcam/ab3760), anti-phosphorylated AMPK (p-AMPK) antibody (CST/2535) and anti-GAPDH antibody (Abcam/ab8245).

Techniques: Activity Assay, Translocation Assay

Journal: Frontiers in Endocrinology

Article Title: Insulin, dibutyryl-cAMP, and glucose modulate expression of patatin-like domain containing protein 7 in cultured human myotubes

doi: 10.3389/fendo.2023.1139303

Figure Lengend Snippet: List of primary antibodies used for immunoblotting.

Article Snippet: p-AMPK Thr 172 , #2535 , Rabbit , 1:1000 , Cell Signaling Technology Inc, USA.

Techniques: Western Blot

Journal: Veterinary Research

Article Title: Curcumin-dependent phenotypic transformation of microglia mediates resistance to pseudorabies-induced encephalitis

doi: 10.1186/s13567-023-01149-x

Figure Lengend Snippet: CUR regulates energy metabolism through AMPK-dependent pathways in PRV-infected BV2 cells. A BV2 cells were infected with/without PRV for 24 h and then treated with/without 20 μM CUR for 24 h. Western blot analysis of p-AMPK Thr172 protein levels (normalized to the t-AMPK protein). β-Actin was used as the loading control ( n = 3). B Relative protein levels of p-AMPK Thr172 . C BV2 cells were treated with different concentrations of Compound C for 24 h, and cell viability was determined by the CCK-8 assay to determine the toxicity range ( n = 4). D BV2 cells were infected with/without PRV or were treated with PRV and an AMPK inhibitor (Compound C or siRNA) alone or in combination for 24 h, followed by treatment with/without 20 μM CUR for 24 h. Western blot analysis of p-AMPK (normalized to the t-AMPK protein), t-AMPK, Gpat4, and LDHa levels (Gpat4 and LDHa values were normalized to the β-actin protein); β-actin was used as the loading control ( n = 3). E Relative p-AMPKThr172 protein levels. F Relative LDHa protein levels. G Relative Gpat4 protein levels. H The extracellular acidification rate of BV2 cells after the different treatments (values normalized to the control) ( n = 4). I The OCR of BV2 cells after the different treatments (values normalized to the control) ( n = 4). J BV2 cell ATP levels in the different treatment groups ( n = 4). K The M1 phenotype-related inflammatory factor TNF-α in BV2 cells was detected using ELISA ( n = 4). L The levels of the M1 phenotype-related inflammatory factor IL-6 in BV2 cells ( n = 4). M The levels of the M2 phenotype-related anti-inflammatory factor IL-4 in BV2 cells ( n = 4). N The levels of the M2 phenotype-related anti-inflammatory factor IL-10 in BV2 cells ( n = 4). All experiments were performed in parallel. The results are presented as the mean ± SD. Statistical significance was determined using one-way ANOVA followed by an LSD post hoc test for multiple comparisons among the groups. * P < 0.05, ** P < 0.01, *** P < 0.001, and NS, not significant.

Article Snippet: Furthermore, the membrane was blocked with 5% nonfat dry milk or 5% BSA in tris-buffered saline containing Tween-20 (TBST) at 25 ℃ and then incubated overnight at 4 °C with the following antibodies: rabbit anti-AMPK (1:2000, bs-1115R, Bioss, China), phosphor(p)-AMPK Thr172 (1:1000, 2531, Cell Signaling, USA), lactate dehydrogenase A (LDHa, 1:1000, WL03271, Wanleibio, China), nuclear factor kappa-B p65 (NF-κB p65, 1:1000, WL01273b, Wanleibio, China), p-NF-κB p65 Ser536 (1:10 000, WL02169, Wanleibio, China), glycerol-3-phosphate acyltransferase 4 (GPAT4, 1:1000, bs-15587R, Bioss, China), Bax (1:10 000, 50,599–2-Ig, Proteintech, China), Caspase-3/p17/p19 (1:1000, 19,677–1-AP, Proteintech, China), Bcl-2 (1:4000, 26,593–1-AP, Proteintech, China), and β-actin (1:5000, 20,536–1-AP, Proteintech, China).

Techniques: Infection, Western Blot, CCK-8 Assay, Enzyme-linked Immunosorbent Assay