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: 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