Journal: Iranian Journal of Basic Medical Sciences

Article Title: Curcumin protects human umbilical vein endothelial cells against high oxidized low density lipoprotein-induced lipotoxicity and modulates autophagy

doi: 10.22038/IJBMS.2021.59969.13297

Figure Lengend Snippet: Curcumin ameliorated autophagy inhibition induced by high ox-LDL in HUVECs through the AMPK/mTOR/p70S6K pathway. A and B: The protein expression levels of LC3-II were measured by Western blotting (n=3). C and D: The protein expression levels of AMPK and p-AMPK were measured by Western blotting (n=3). E and F: The protein expression levels of mTOR and p-mTOR were measured by Western blotting (n=3). G and H: The protein expression levels of p70S6K and p-p70S6K were measured by Western blotting. Expression of these proteins was quantified by densitometry using ImageJ software (n=3). The data are shown as the means±SDs (* P < 0.01 vs. the control group; ** P < 0.01 vs. the control group; # P <0.05 vs. the high ox-LDL group; ## P < 0.01 vs. the high ox-LDL group; & P < 0.05 vs. the compound C group). AMPK, AMP-activated protein kinase; mTOR, mammalian target of rapamycin; p70S6K, p70 ribosomal S6 protein kinase

Article Snippet: Protein extracts were isolated, separated on a 12.5% SDS-PAGE gel and transferred onto a 0.22 μm PVDF membrane (Epizyme), which was incubated with the following primary antibodies: peroxisome proliferator-activated receptor γ (PPARγ) (16001-1-AP, Proteintech, Wuhan, China), interleukin-6 (IL-6) (ab233706, Abcam, Cambridge, 1:1000), interleukin-10 (IL-10) (ab133575, Abcam, 1:5000), tumor necrosis factor alpha (TNF-α) (17590-1-AP, Proteintech, Wuhan, China), microtubule-associated protein 1 light chain 3 (LC3) (2775S, Cell Signaling Technology, MA, USA, 1:1000), AMPK (HN0506, HuaBio, Hangzhou, China, 1:1000), mTOR (HN0824, HuaBio, 1:500), p70S6K (HJ0505, HuaBio, 1:1000), phosphorylated AMPK (p-AMPK) (bs-8813R, Bioss, Beijing, China, 1:1000), phosphorylated mTOR (p-mTOR) (2971S, Cell Signaling Technology, 1:500), and phosphorylated p70S6K (p-p70S6K) (9234S, Cell Signaling Technology, 1:1000). β-Actin (AC026, ABclonal, Wuhan, China, 1:50000) was used as an internal control.

Techniques: Inhibition, Expressing, Western Blot, Software

Journal: Viruses

Article Title: The Function behind the Relation between Lipid Metabolism and Vimentin on H9N2 Subtype AIV Replication.

doi: 10.3390/v14081814

Figure Lengend Snippet: Figure 4. The cellular AMPK expression and phosphorylation level. AMPK protein expressions and phosphorylation in HeLa and KO HeLa cells with or without viral infection were detected with Western blotting. (A) The expression and phosphorylation of AMPK protein on KO HeLa cells compared with HeLa cells. (B) The expression and phosphorylation of AMPK protein on HeLa cells after viral infection. (C) The expression and phosphorylation of AMPK protein on KO HeLa cells after viral infection.

Article Snippet: GAPDH monoclonal antibody was purchased from Enogene Biologicals (Nanjing, China). β-actin monoclonal antibody, AMPK monoclonal antibody and AMPK phosphorylated monoclonal antibody P-AMPK were purchased from BOSTER Biologicals (Nanjing, China).

Techniques: Expressing, Phospho-proteomics, Infection, Western Blot

Journal: Viruses

Article Title: The Function behind the Relation between Lipid Metabolism and Vimentin on H9N2 Subtype AIV Replication.

doi: 10.3390/v14081814

Figure Lengend Snippet: Figure 7. Model of lipid metabolism and vimentin on H9N2 subtype AIV. When vimentin was knocked down, the phosphorylation level of AMPK was decreased, resulting in the decreased level of HMGCR phosphorylation, which increased the enzyme activity, thereby increasing cholesterol. After the destruction of lipid rafts, the virus binding was inhibited, and cholesterol helped to stabilize the structure of lipid rafts, which might help the virus bind to cells.

Article Snippet: GAPDH monoclonal antibody was purchased from Enogene Biologicals (Nanjing, China). β-actin monoclonal antibody, AMPK monoclonal antibody and AMPK phosphorylated monoclonal antibody P-AMPK were purchased from BOSTER Biologicals (Nanjing, China).

Techniques: Phospho-proteomics, Activity Assay, Virus, Binding Assay

Journal: Scientific reports

Article Title: Features of an altered AMPK metabolic pathway in Gilbert's Syndrome, and its role in metabolic health.

doi: 10.1038/srep30051

Figure Lengend Snippet: Figure 1. Comparison of measures of the AMPK pathway between the study groups (GS, C), including all subjects (male and female). Levels of (phosphorylated) proteins (AMPK α1/α2, Ppar α and γ, PgC 1α) were analysed using the method of flow cytometry. Data are expressed as relative fluorescence units [rfU], and compared between subjects with Gilbert’s syndrome (GS; UGT1A1*28 promoter mutation), and controls (C). * Indicates significant differences between groups (p ≤ 0.05). Medians can be found in Table 2. Abbreviations: pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1.

Article Snippet: The antibody set-up used was as follows: anti-phospho-AMPK: rabbit anti-human monoclonal to AMPK α1 (phos-T183) and AMPK α2 (phos-T172), (ab133448, Abcam); secondary antibody to phospho-AMPK: goat anti-rabbit IgG H & L AlexaFluor 488, (ab150077, Abcam); anti-PgC 1α: rabbit anti-human polyclonal to PgC 1α, PE-labelled, (orb124814, Biorbyt); anti-phospho-Ppar α: rabbit anti-human polyclonal to Ppar α (phos-Ser12), FITC-labelled, (bs-4055R-FITC, Bioss); anti-phospho-Ppar γ: rabbit anti-human polyclonal to Ppar γ (phos-Ser112), AlexaFluor 647 labelled, (bs-3737R-A647, Bioss).

Techniques: Comparison, Flow Cytometry, Fluorescence, Mutagenesis

Journal: Scientific reports

Article Title: Features of an altered AMPK metabolic pathway in Gilbert's Syndrome, and its role in metabolic health.

doi: 10.1038/srep30051

Figure Lengend Snippet: Figure 3. Correlations of UCB (a–d) and the UGT1A1 genotype (e–h), with measures of the AMPK pathway. Figure 3 illustrates gender-specific correlations of UCB and the UGT1A1 genotype with measures of the AMPK pathway. Bivariate correlations between UCB/UGT1A1 genotype (-TA repeats: 6/6 controls, 6/7 heterozygous, 7/7 Gilbert’s syndrome) and measures of the AMPK pathway were calculated for each gender (m = male, f = female), using the model of Spearman’s rho. R coefficients and p-values (p ≤ 0.05; in brackets) are as follows: UCB * pAMPK α1/α2: m 0.594 (0.000); f 0.255 (0.122), UCB * PgC1 α: m 0.376 (0.001); f 0.467 (0.003), UCB * pPpar α: m 0.435 (0.000); f 0.575 (0.000), UCB * pPpar γ: m 0.354 (0.001); f 0.324 (0.047), UGT1A1 * pAMPK α1/α2: m 0.541 (p 0.000); f 0.156 (p 0.362), UGT1A1 * PgC1 α: m 0.265 (p 0.023); f 0.551 (p 0.001), UGT1A1 * Ppar α: m 0.365 (p 0.002); f 0.661 (p 0.000), UGT1A1 * Ppar γ: m 0.191 (p 0.023); f 0.435 (p 0.008). Abbreviations: UCB: unconjugated bilirubin; pAMPK α1/α2: Phosphorylated 5′-AMP activated kinase; pPpar α: Phosphorylated peroxisome proliferator activated receptor alpha; pPpar γ: Phosphorylated peroxisome proliferator activated receptor gamma; PgC 1α: Peroxisome proliferator-activated receptor c coactivator 1; WT: wild type (control subjects); GS: Gilbert’s syndrome.

Article Snippet: The antibody set-up used was as follows: anti-phospho-AMPK: rabbit anti-human monoclonal to AMPK α1 (phos-T183) and AMPK α2 (phos-T172), (ab133448, Abcam); secondary antibody to phospho-AMPK: goat anti-rabbit IgG H & L AlexaFluor 488, (ab150077, Abcam); anti-PgC 1α: rabbit anti-human polyclonal to PgC 1α, PE-labelled, (orb124814, Biorbyt); anti-phospho-Ppar α: rabbit anti-human polyclonal to Ppar α (phos-Ser12), FITC-labelled, (bs-4055R-FITC, Bioss); anti-phospho-Ppar γ: rabbit anti-human polyclonal to Ppar γ (phos-Ser112), AlexaFluor 647 labelled, (bs-3737R-A647, Bioss).

Techniques: Control