Journal: NPJ Science of Food

Article Title: A soluble garlic polysaccharide supplement alleviates fatigue in mice

doi: 10.1038/s41538-024-00340-4

Figure Lengend Snippet: AMPK, P-AMPK, and PGC-1α protein expression in muscle.

Article Snippet: The antibodies against AMPK (#GB112669), p-AMPK (#GB114323), PGC-1α (#GB11912), GAPDH (#GB15002), AMPK (#GB23303), p-AMPK (#GB23303), PGC-1α (#GB23303) and GAPDH (#GB23301) were purchased from Servicebio (Wuhan, China).

Techniques: Expressing

Journal: Neural Regeneration Research

Article Title: Bradykinin postconditioning protects rat hippocampal neurons after restoration of spontaneous circulation following cardiac arrest via activation of the AMPK/mTOR signaling pathway

doi: 10.4103/1673-5374.337049

Figure Lengend Snippet: Effect of bradykinin postconditioning on the immunopositivity of AMPK/mTOR signaling pathway, autophagy-related proteins, and brain injury marker in rats with restoration of spontaneous circulation, as detected by immunohistochemistry. Compared with the Sham group, p-mTOR and p62 immunopositivity in the ROSC group was significantly decreased, S100β, p-AMPK, and LC3 immunopositivity was significantly increased, and NBR1 immunopositivity remained unchanged. Compared with the ROSC group, p-mTOR, p62, and S100β immunopositivity in the BK group was significantly decreased, while p-AMPK, NBR1, and LC3 immunopositivity was significantly increased. Compared with the BK group, LC3, p-AMPK, and NBR1 immunopositivity in the CP+BK group was significantly decreased, whereas S100β, p62, and p-mTOR immunopositivity was significantly increased; LC3, p-AMPK, and NBR1 immunopositivity in the Ra+BK group was significantly increased, while S100β, p62, and p-mTOR immunopositivity was significantly decreased. Scale bars: 50 μm. All data are presented as the mean ± SD ( n = 8). * P < 0.05, ** P < 0.01, vs . Sham group; # P < 0.05, ## P < 0.01, vs . ROSC group; † P < 0.05, †† P < 0.01, vs . BK group (one-way analysis of variance followed by Tukey’s post hoc test). BK: Bradykinin; CP: compound C; IOD: integrated optical density; LC3: microtubule-associated protein 1 light chain 3; NBR1: neighbor of breast cancer 1 gene; p-AMPK: phosphorylated adenosine-monophosphate activated protein kinase; p-mTOR: p-mechanistic target of rapamycin; Ra: rapamycin; ROSC: restoration of spontaneous circulation; S100β: S100 calcium-binding protein B; Sham: sham operation.

Article Snippet: The diluted primary antibodies [S100 calcium-binding protein B (S100β, a brain injury marker; 1:100; rabbit; Cat# ab52642, RRID: AB_882426; Abcam Co., Cambridge, MA, USA), phosphorylated mTOR (p-mTOR; 1:100; rabbit; Cat# ab109268, RRID: AB_10888105; Abcam Co.), neighbor of breast cancer 1 gene (NBR1; an autophagy-related protein; 1:100; rabbit; Cat# DF12049, RRID: AB_2844854; Abcam Co.), microtubule-associated protein 1 light chain 3 (LC3) I/II (an autophagy-related protein; 1:100; rabbit; Cat# AF5402, RRID: AB_2837886; Affinity Biosciences, Cincinnati, OH, USA), phosphorylated AMPK (p-AMPK; 1:100; rabbit; Cat# AF3423, RRID: AB_2834865; Affinity Biosciences), and p62 (an autophagy-related protein; 1:100; rabbit; Cat# 18420-1-AP, RRID: AB_10694431; ProteinTech Group, Inc., Rosemont, IL, USA)] were then added, and the sections were incubated in a wet box overnight at 4°C.

Techniques: Marker, Immunohistochemistry, Binding Assay

Journal: Neural Regeneration Research

Article Title: Bradykinin postconditioning protects rat hippocampal neurons after restoration of spontaneous circulation following cardiac arrest via activation of the AMPK/mTOR signaling pathway

doi: 10.4103/1673-5374.337049

Figure Lengend Snippet: Effect of bradykinin postconditioning on the AMPK/mTOR signaling pathway and apoptosis-related protein expression in rats with restoration of spontaneous circulation, as detected by western blot. Original images of bands were shown in Additional Figure 2 . All data are presented as the mean ± SD ( n = 8). ** P < 0.01, vs . Sham group; ## P < 0.01, vs . ROSC group; †† P < 0.01, vs . BK group (one-way analysis of variance followed by Tukey’s post hoc test). BK: Bradykinin; caspase-3: cysteine-aspartic acid proteases; CP: compound C; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; p-AMPK: phosphorylated adenosine monophosphate-activated protein kinase; p-mTOR: phosphorylated mechanistic target of rapamycin; Ra: rapamycin: ROSC: restoration of spontaneous circulation; Sham: sham operation.

Article Snippet: The diluted primary antibodies [S100 calcium-binding protein B (S100β, a brain injury marker; 1:100; rabbit; Cat# ab52642, RRID: AB_882426; Abcam Co., Cambridge, MA, USA), phosphorylated mTOR (p-mTOR; 1:100; rabbit; Cat# ab109268, RRID: AB_10888105; Abcam Co.), neighbor of breast cancer 1 gene (NBR1; an autophagy-related protein; 1:100; rabbit; Cat# DF12049, RRID: AB_2844854; Abcam Co.), microtubule-associated protein 1 light chain 3 (LC3) I/II (an autophagy-related protein; 1:100; rabbit; Cat# AF5402, RRID: AB_2837886; Affinity Biosciences, Cincinnati, OH, USA), phosphorylated AMPK (p-AMPK; 1:100; rabbit; Cat# AF3423, RRID: AB_2834865; Affinity Biosciences), and p62 (an autophagy-related protein; 1:100; rabbit; Cat# 18420-1-AP, RRID: AB_10694431; ProteinTech Group, Inc., Rosemont, IL, USA)] were then added, and the sections were incubated in a wet box overnight at 4°C.

Techniques: Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Dihydromyricetin Ameliorates Inflammation-Induced Insulin Resistance via Phospholipase C-CaMKK-AMPK Signal Pathway

doi: 10.1155/2021/8542809

Figure Lengend Snippet: The details of antibodies used in this study.

Article Snippet: p-AMPK , Polyclonal , Abbkine , ABP50452 , 1 : 1000.

Techniques:

Journal: Nutrients

Article Title: 1,25-Dihydroxyvitamin D 3 Provides Benefits in Vitiligo Based on Modulation of CD8+ T Cell Glycolysis and Function

doi: 10.3390/nu15214697

Figure Lengend Snippet: Correlation between 1,25(OH)₂D₃ and immune cells. ( a – c ). The relationship between serum 1,25(OH)₂D₃ levels in patients with different immunological data was evaluated and revealed a significant negative correlation between serum 1,25(OH)₂D₃ and the counts of T cells and frequency of CD8+ T cells/ T cells and IFN+ CD8+ T cells/ CD8+ T cells. ( d ). Volcano plot of detectable mRNA profiles in participants receiving vitamin D ( n = 47) or placebo ( n = 47). Red and blue plots represent aberrantly expressed mRNAs with p < 0.05 and absolute log2FC > 2, and grey plots indicate transcripts not significantly changed. The abscissa means the value of fold change in mRNA expression. The ordinate shows the −log10 of the p -value for each mRNA, representing the strength of the association. ( e ). A plot heatmap showing the top 50 gene expression profiles of differentially expressed genes (DEGs). The magnitude of change for each variable is indicated by the color gradient. The most intense shades of red in the red gradient represent the greatest magnitudes of change for the dependent variable. As the color gradient becomes whiter, it indicates no change in the dependent variable. As the color gradient flows from white to more intense shades of blue, it indicates a decrease in the values of the dependent variable. ( f ) The KEGG pathway enrichment analysis revealed that DEGs involved in the glycolysis/gluconeogenesis and AMPK signaling pathways were enriched significantly in the participants receiving vitamin D. AMPK, AMP-activated protein kinase.

Article Snippet: The membrane was blocked in 5% bovine serum albumin for 1 h and then incubated at 4 °C overnight with primary antibodies, including AMPK (1:1000, ABclonal, Wuhan, China) and p -AMPK (1:1000, ABclonal, Wuhan, China). β-actin (1:2000, ABclonal, Wuhan, China) was used as an internal reference.

Techniques: Expressing, Gene Expression, Protein-Protein interactions

Journal: Nutrients

Article Title: 1,25-Dihydroxyvitamin D 3 Provides Benefits in Vitiligo Based on Modulation of CD8+ T Cell Glycolysis and Function

doi: 10.3390/nu15214697

Figure Lengend Snippet: Regulation of CD8+ T cell glucose metabolism and immunity by 1,25(OH)₂D₃ via AMPK signaling pathway. ( a ). The uptake of 2-NBDG by TALL-104 cells in each group was observed using fluorescence assay. ( b ). Fluorescence intensity of 2-NBDG uptake by TALL-104 cells in each group. ( c ). LDH activity of TALL-104 cells in each group. ( d ). HK activity of TALL-104 cells in each group. ( e ). Levels of IFN-γ in the supernatant of TALL-104 cells in each group. ( f ). GLUT1, LDHA, HKII and IFNG mRNA expression in each group. ( g ). The expression and phosphorylation of AMPK in each group. ( h ). The expression and phosphorylation of AMPK were analyzed using Western blotting after treatment of TALL-104 cells with 1,25(OH)₂D₃ and/or Compound C. ( i ). 2-NBDG fluorescence intensity, LDH activity, HK activity and IFN-γ level in supernatant of TALL-104 cells treated with 1,25(OH)₂D₃ and/or Compound C. Data are from a representative one of three experiments and are expressed as mean ± standard deviation (SD). Individual data points are represented as black dots. *, p < 0.05; **, p < 0.01; ***, p < 0.001; 2-NBDG, 2-NBD-Glucose; LDH, lactic dehydrogenase; HK, hexokinase; AMPK, AMP-activated protein kinase; CC, Compound C; VD, 1,25(OH)₂D₃.

Article Snippet: The membrane was blocked in 5% bovine serum albumin for 1 h and then incubated at 4 °C overnight with primary antibodies, including AMPK (1:1000, ABclonal, Wuhan, China) and p -AMPK (1:1000, ABclonal, Wuhan, China). β-actin (1:2000, ABclonal, Wuhan, China) was used as an internal reference.

Techniques: Fluorescence, Activity Assay, Expressing, Phospho-proteomics, Western Blot, Standard Deviation

Journal: Nutrients

Article Title: 1,25-Dihydroxyvitamin D 3 Provides Benefits in Vitiligo Based on Modulation of CD8+ T Cell Glycolysis and Function

doi: 10.3390/nu15214697

Figure Lengend Snippet: 1,25(OH)₂D₃ supplementation can lead to CD8+ T cell dysfunction, resulting in skin repigmentation. Via a supply of exogenous 1,25(OH)₂D₃, the phosphorylation level of AMPK in CD8+ T cells is upregulated, which leads to reduced levels of glycolysis, as evidenced by the downregulation of glucose uptake and activities of the key enzymes of glycolysis, HK and LDH. These changes altered the effect of CD8+ T cells, leading to a decrease in proliferation and IFN-γ secretion of CD8+ T cells, consequently decreasing their cytotoxicity against melanocytes. As described above, the immunomodulatory effect of 1,25(OH)₂D₃ on CD8+ T cells causes skin repigmentation, which we have verified in the MAV mouse model. LDH, lactic dehydrogenase; HK, hexokinase; AMPK, AMP-activated protein kinase.

Article Snippet: The membrane was blocked in 5% bovine serum albumin for 1 h and then incubated at 4 °C overnight with primary antibodies, including AMPK (1:1000, ABclonal, Wuhan, China) and p -AMPK (1:1000, ABclonal, Wuhan, China). β-actin (1:2000, ABclonal, Wuhan, China) was used as an internal reference.

Techniques: Phospho-proteomics