Journal: The Journal of international medical research

Article Title: The therapeutic potential of irisin in alleviating acute lung injury via inflammation and ferroptosis modulation.

doi: 10.1177/03000605251340338

Figure Lengend Snippet: Figure 6. Western blotting analysis on the effects of irisin on the expression level of ferroptosis-associated proteins. (a) Representative western blotting images for the expression of ACSL4, COX-2, GPX4, p-AMPK, and t-AMPK in lung tissues. GAPDH was selected as the loading control protein. (b) Quantification analysis of the related bands of ACSL4, COX-2, GPX4, p-AMPK, and t-AMPK in lung tissues. Lung tissues were harvested on day 3 post-CLP. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post-hoc test. n ¼ 6 per group. Data are presented as means SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: Subsequently, the membranes were incubated with rabbit anti-AMPK (MCE, Cat# HY-P80541), anti-p-AMPK (MCE, HYP80452), GPX4 (MCE, HY-P80450), ACSL4 (Santa Cruz Biotechnology, Dallas, Texas, USA; Cat# sc-365230), COX-2 (Abcam, Cambridge, United Kingdom; Cat# ab283574), and GAPDH (MCE, HY-P80137) antibodies at 4 C overnight.

Techniques: Western Blot, Expressing, Control

Journal: BMB Reports

Article Title: Corin inhibits microglial inflammatory activation by suppressing mitochondrial dysfunction in intracerebral hemorrhage

doi: 10.5483/BMBRep.2025-0175

Figure Lengend Snippet: Effects of corin overexpression on OGD/H-induced mitochondrial dysfunction in HAPI microglia. Following 24 h pre-transduction with corin overexpression or blank lentivirus, cells underwent 24 h OGD/H exposure. Measurements included (A) OCR, (B-F) Mitochondrial complex activities (I, II, III, IV, V), (G) ATP generation, (H) p-AMPK/AMPK protein levels. Data presented as mean ± SD (n = 3). All experiments were repeated three times. Significant differences between groups were assessed using one-way ANOVA with Tukey’s post-hoc test. ***P < 0.001 vs. control. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. OGD/H＋vector.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies targeting Corin (Proteintech, 83566-1-RR), Iba-1 (Proteintech, 81728-1-RR), phosphorylated AMPK (p-AMPK; Abclonal, AP1002), AMPK (Proteintech, 10929-2-AP), and β-actin (Proteintech, 66009-1-Ig).

Techniques: Over Expression, Transduction, Control

Journal: BMB Reports

Article Title: Corin inhibits microglial inflammatory activation by suppressing mitochondrial dysfunction in intracerebral hemorrhage

doi: 10.5483/BMBRep.2025-0175

Figure Lengend Snippet: Overexpression of corin mitigates OGD/H-induced cell migration impairment, inflammatory response, and mitochondrial dysfunction in HAPI microglia via the AMPK pathway. Cells were pre-transduced with corin-overexpressing or control lentivirus for 24 h, followed by OGD/H and treatment with 10-μM Compound C for 24 h. Subsequent assessments included: (A) cell viability, (B) migration, (C) iNOS and Arg-1 mRNA levels, (D) Iba-1, p-AMPK, and AMPK protein levels, (E) OCR, (F-J) activities of mitochondrial complexes I-V, and (K) ATP generation. Data presented as mean ± SD (n = 3). All experiments were repeated three times. Significant differences between groups were assessed using one-way ANOVA with Tukey’s post-hoc test. *P < 0.05, **P < 0.01, ***P < 0.001 vs. OGD/H＋vector＋vehicle. # P < 0.05, ## P < 0.01, ### P < 0.001 vs. OGD/H＋Corin+vehicle.

Article Snippet: Membranes were incubated overnight at 4°C with primary antibodies targeting Corin (Proteintech, 83566-1-RR), Iba-1 (Proteintech, 81728-1-RR), phosphorylated AMPK (p-AMPK; Abclonal, AP1002), AMPK (Proteintech, 10929-2-AP), and β-actin (Proteintech, 66009-1-Ig).

Techniques: Over Expression, Migration, Transduction, Control

Journal: Chinese Herbal Medicines

Article Title: Ganoderic acid a derivative induces apoptosis of cervical cancer cells by inhibiting JNK pathway

doi: 10.1016/j.chmed.2024.07.002

Figure Lengend Snippet: (A) Target prediction analysis results. (B) Prediction of GaAD19-JNK interaction. (C–E) Western blot analysis of HeLa cells with GaAD19 treatment for 24 h on expression of JNK, p-JNK, ERK, p-ERK, p38, p-p38 (C), AMPK α 1, p-AMPK α 1-S485, p-AMPK α 1-S496, AMPK β 1, p-AMPK β 1-S108, Akt, p-Akt (D), MAP2K4, MAP2K7 (E), and other proteins (means ± SD, n = 3). (F) Effect of GaAD19 on mRNA level of JNK pathway membrane protein receptor in HeLa cells was determined by qPCR (means ± SD, n = 3). ( ** P < 0.01, *** P < 0.001 vs control group, ns indicates no significant difference).

Article Snippet: p-AMPK β 1-S108 , ABclonal , AP0597 , 1:200.

Techniques: Western Blot, Expressing, Membrane, Control

Journal: Metabolism: clinical and experimental

Article Title: Serum metabolomics analysis reveals increased lipid catabolism in mildly hyperbilirubinemic Gilbert's syndrome individuals.

doi: 10.1016/j.metabol.2021.154913

Figure Lengend Snippet: Fig. 4. AMPK, PPARα, PGC1α and lipid catabolism. The figure presents the % increase of AMPK, PPARα and PGC-1α measured in PBMCs of GS vs. control individuals, as previously published by Mölzer et al. [22] and correlation coefficients (r) between UCB, AMPK; PPARα and PGC-1 α. The table shows correlations coefficients and p-values of AMPK (blue), PGC1α (orange), PPARα (green) and UCB (yellow) with their downstream lipid metabolites in the whole study population using Pearson or Spearman-Rho. Significant correlations are in bold type, t = trend.

Article Snippet: The following antibody set-up was used: rabbit anti-human monoclonal to AMPK α1 (phos-T183) and AMPKα2 (phos-T172) (ab133448, Abcam) and secondary antibody: goat anti-rabbit IgG H & L AlexaFluor 488 (ab150077, Abcam); rabbit anti-human polyclonal to PgC1α PE-labelled (orb124814, Biorbyt) and rabbit anti-human polyclonal to PPARα (phos-Ser12) FITClabelled (bs-4055R-FITC, Bioss).

Techniques: Control

Journal: Molecular Metabolism

Article Title: Exercise-induced specialized proresolving mediators stimulate AMPK phosphorylation to promote mitochondrial respiration in macrophages

doi: 10.1016/j.molmet.2022.101637

Figure Lengend Snippet: SPMs stimulate AMPKα phosphorylation to enhance mitochondrial metabolism in macrophages. ( A ) Representative immunoblot and quantification (right) of phosphorylated AMPKα (Thr172) and total AMPK in BMDMs treated without or with 1 nM RvD1 for 1h. ( B – E ) BMDMs treated without or with 500 nM Compound-C (CompC) and 1 nM of ( B )RvD1, ( C ) RvD2, ( D ) RvE1, or ( E ) MaR1 for 1h. After treatment, cells were subjected to a mitochondrial stress assay and OCR values assessed (top panels B – E ) and derived mitochondrial respiratory parameters were quantified (below). Data expressed as mean ± SEM; n = 3 ( A ) or n = 3–6 ( B – E ); ∗ P < 0.05, ∗∗ P < 0.01; two-tailed Student's t -test (a) or Two-way ANOVA with Holm-Šídák post-test ( B – E ).

Article Snippet: Cells were then stained with a polyclonal, rabbit anti-mouse FITC-labeled p-AMPKα1(Thr172) antibody (Biorbyt, cat. # orb8540) in a 100 μL staining volume (perm/wash buffer) and incubated 30 min on ice.

Techniques: Western Blot, Derivative Assay, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Exercise-induced specialized proresolving mediators stimulate AMPK phosphorylation to promote mitochondrial respiration in macrophages

doi: 10.1016/j.molmet.2022.101637

Figure Lengend Snippet: 5-lipoxygenase derived lipid mediators contribute to exercise-induced mitochondrial function via AMPK activation. ( A ) Flow cytometric analysis (MFI) of intracellular AMPKα1(Thr172) phosphorylation in PMs isolated from WT and Alox5 −/− mice following exercise. ( B ) FlowJo V10.8.1 generated t -SNE map ( B,C ) depicted in a 2D dimensionally reduced dataspace showing all events combined from each experimental groups. The color axis overlay in ( B ) depicts metacluster populations identified using FlowSOM, an unbiased machine learning clustering algorithm. ( C ) The percent frequency of each FlowSOM population (highlighted in red) for all samples combined, is denoted atop of the respective t -SNE map. ( D ) The fluorescence distribution of each surface marker for the respective FlowSOM metacluster population is shown in red (vs the distribution for all cells shown in grey) in the histogram overlays. ( E ) MFI of TMRM for each FlowSOM metacluster population from WT and Alox5 −/− mice following exercise. Data expressed as mean ± SEM; n = 5 ( A and E ); ∗ P < 0.05; two-tailed Student's t -test.

Article Snippet: Cells were then stained with a polyclonal, rabbit anti-mouse FITC-labeled p-AMPKα1(Thr172) antibody (Biorbyt, cat. # orb8540) in a 100 μL staining volume (perm/wash buffer) and incubated 30 min on ice.

Techniques: Derivative Assay, Activation Assay, Isolation, Generated, Fluorescence, Marker, Two Tailed Test