ampk inhibition  (MedChemExpress)

Journal: Bioactive Materials

Article Title: Melatonin-incorporated brain extracellular matrix hydrogel enhances NSCs mitochondrial metabolism to promote neuroregeneration via the AMPK-PGC-1α-NRF1/TFAM axis after spinal cord injury

doi: 10.1016/j.bioactmat.2026.04.006

Figure Lengend Snippet: Melatonin activates AMPK signaling and enhances mitochondrial function in vitro. (A) GO enrichment bar plot of differentially expressed genes (DEGs) between Control and Melatonin-treated NSCs. (B) KEGG pathway enrichment bar plot of DEGs between Control and Melatonin groups. (C) Heatmap of selected DEGs associated with neuronal differentiation and mitochondrial function. DEGs were defined as transcripts with FDR <0.05. (D) Representative Western blots showing phosphorylated AMPK (p-AMPK, Thr172) and phosphorylated ACC (p-ACC, Ser79) in Control, Melatonin, Inhibitor, and Melatonin + Inhibitor groups. (E) Densitometric analysis of p-AMPK/total AMPK and p-ACC/GAPDH ratios. (F) RT-qPCR analysis of Ppargc1a and Tfam expression, normalized to GAPDH and presented as fold change relative to the Control group. (G) Representative Western blots of mitochondrial oxidative phosphorylation (OXPHOS) complexes I-V. (H) Densitometric quantification of OXPHOS complexes I-V, normalized to GAPDH (or the corresponding loading control). (I) Representative JC-1 fluorescence images indicating mitochondrial membrane potential (ΔΨm). (J) Quantification of the red/green JC-1 fluorescence ratio from (I). (K) Schematic representation of the proposed melatonin-AMPK-ACC-PGC-1α-NRF1/TFAM signaling axis driving mitochondrial biogenesis in NSCs. All quantitative data (E, F, H, J) are presented as mean ± SD. Statistical significance was assessed using one-way ANOVA followed by Holm–Sidak's multiple comparisons test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. K created with BioRender.com .

Article Snippet: For AMPK inhibition experiments, BAY-3827 (HY-112083, MedChemExpress, USA), a selective AMPK inhibitor, was used at a final concentration of 2 μM for 24 h. The mitochondrial membrane potential was measured using the JC-1 Mitochondrial Membrane Potential Assay Kit (C2003S, Beyotime Biotechnology, China).

Techniques: In Vitro, Control, Western Blot, Quantitative RT-PCR, Expressing, Phospho-proteomics, Fluorescence, Membrane

Journal: Bioactive Materials

Article Title: Melatonin-incorporated brain extracellular matrix hydrogel enhances NSCs mitochondrial metabolism to promote neuroregeneration via the AMPK-PGC-1α-NRF1/TFAM axis after spinal cord injury

doi: 10.1016/j.bioactmat.2026.04.006

Figure Lengend Snippet: Molecular validation of neural repair and mechanism activation in spinal cord tissue. Western blot and qPCR analyses of spinal cord tissue lysates from Sham, SCI, BEM, NSCs@BEM, and NSCs@MT/BEM groups. (A) Representative Western blots for the neuronal marker TUJ1 and the glial scar marker GFAP. (B) Representative Western blots for phosphorylated AMPK (p-AMPK), phosphorylated ACC (p-ACC), and their respective total proteins. (C) Representative Western blots for the five oxidative phosphorylation (OXPHOS) complex subunits. (D) Densitometric quantification of TUJ1 and GFAP protein levels. (E) Densitometric quantification of the p-AMPK/total AMPK and p-ACC/total ACC ratios. (F) Densitometric quantification of OXPHOS complex protein levels. (G) Relative mRNA expression of neural markers (TUJ1, GFAP, Olig2) and key mitochondrial biogenesis regulators (Ppargc1a, Tfam) determined by qPCR. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA with Holm–Sidak's multiple comparisons test. (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: For AMPK inhibition experiments, BAY-3827 (HY-112083, MedChemExpress, USA), a selective AMPK inhibitor, was used at a final concentration of 2 μM for 24 h. The mitochondrial membrane potential was measured using the JC-1 Mitochondrial Membrane Potential Assay Kit (C2003S, Beyotime Biotechnology, China).

Techniques: Biomarker Discovery, Activation Assay, Western Blot, Marker, Phospho-proteomics, Expressing

Journal: iScience

Article Title: Disulfidptosis in PCOS pathogenesis: Multi-omics identification of LRPPRC as a diagnostic biomarker and therapeutic target

doi: 10.1016/j.isci.2026.116561

Figure Lengend Snippet: Synergistic induction of disulfidptosis in granulosa cells by LRPPRC deficiency and pharmacological GLUT1 inhibition (A) Representative F-actin (red) and DAPI (blue) fluorescence staining images of KGN cells. Cells with control knockdown (Sh NC) or LRPPRC knockdown (Sh LRPPRC) were treated with DMSO (vehicle control) or the GLUT1-specific inhibitor BAY-876 (BAY), and subsequently cultured under glucose-sufficient (+Glc), glucose-deprivation (-Glc), or 2-mercaptoethanol-rescued (2ME/-Glc) conditions. The severe actin cytoskeletal collapse induced by the combination of LRPPRC deficiency, BAY-876, and glucose starvation is completely rescued by the disulfide-reducing agent 2 ME. Images are representative of n = 3 independent biological replicates. Scale bars, 50 μm. (B) Flow cytometry analysis quantifying cell death. Upper and middle panels display representative flow histograms showing the percentage of dead cells (PI-positive population) for the indicated treatment groups under +Glc and -Glc conditions, respectively. The lower panel is a quantitative bar graph summarizing the PI-positive percentage. The results demonstrate a significant synergistic lethal effect when combining LRPPRC knockdown with BAY-876 treatment under glucose deprivation. Data are presented as the mean ± SD of n = 3 independent biological replicates. ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 for the indicated pairwise comparisons.

Article Snippet: BAY-876 , MedChemExpress , Cat# HY-100017.

Techniques: Inhibition, Fluorescence, Staining, Control, Knockdown, Cell Culture, Flow Cytometry

Journal: Bone & Joint Research

Article Title: Piezo1 drives fibroblast activation in epidural fibrotic remodelling via the ET-1/HIF-1α pathway

doi: 10.1302/2046-3758.158.BJR-2025-0662.R1

Figure Lengend Snippet: The hypoxia-inducible factor 1-alpha (HIF-1α)/transforming growth factor beta 1 (TGF-β1)/Smad3 pathway functions downstream of Piezo1 in the activation of fibroblasts. a) Western blot analysis of NIH/3T3 fibroblasts treated with DMSO or Yoda1 (n = 3). b) Western blot analysis of NIH/3T3 fibroblasts treated with Yoda1 and the HIF-1α inhibitor BAY 87-2243 (n = 3). c) and d) Western blot analysis of NIH/3T3 fibroblasts cultured with type I rat tail collagen and the HIF-1α inhibitor BAY 87-2243 (n = 3). e) and f) Western blot analysis of NIH/3T3 fibroblasts cultured with type I rat tail collagen and the TGF-β1 inhibitor SB-431542 (n = 3). g) Western blot analysis of NIH/3T3 fibroblasts cultured with type I rat tail collagen and the Smad3 inhibitor SIS 3 (n = 3). h) and i) Western blot analysis of NIH/3T3 fibroblasts cultured with type I rat tail collagen and the Piezo1 inhibitor GsMTX4 (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, independent-samples t -test (two groups) or analysis of variance (multiple groups).

Article Snippet: Alternatively, the following drugs were used to stimulate NIH/3T3 cells: Yoda1 (25 μM; HY-P1410), a chemical activator of Piezo1; BAY 87-2243 (1 μM; HY-15836), an inhibitor of hypoxia-inducible factor 1-alpha (HIF-1α); SB-431542 (10 μM; HY-10431), an inhibitor of TGF-β1; SIS 3 (10 μM; HY-13013), an inhibitor of Smad3; GsMTX4 (2.5 μM; HY-P1410); Nimodipine (60 μM; HY-B0265), a calcium channel blocker; and Bosentan (1 μM; HY-A0013, all MedChemExpress), an endothelin-1 (ET-1) receptor antagonist.

Techniques: Activation Assay, Western Blot, Cell Culture

Journal: CNS Neuroscience & Therapeutics

Article Title: Recombinant Artemin‐Fc Fusion Protein Attenuates TLR4 / NF ‐ κB ‐Associated Neuroinflammation and Modulates Inhibitory/Excitatory Synaptic Marker Expression After Spinal Cord Injury

doi: 10.1002/cns.71086

Figure Lengend Snippet: Effect of ARTN on the nuclear translocation of p‐NF‐κB p65 in LPS‐stimulated BV‐2 cells. (A) BV‐2 cells were stimulated with LPS (1 μg/mL) in the absence or presence of ARTN for 24 h, followed by detection of the p‐NF‐kB p65 subunit translocation by immunofluorescence. p‐NF‐κB p65 is shown in green, blue fluorescence represents the nuclear marker DAPI. Scale bar = 50 μm. (A boxed region illustrates a representative region with high power images, scale bar = 5 μm) (B) BV‐2 cells were stimulated with LPS (1 μg/ml) in the absence or presence of ARTN for 24 h, and TLR4, p‐NF‐κB p65 and NF‐κB p65 levels were determined by Western blot. β‐actin were used as endogenous controls. (C, D) Quantitative analysis of TLR4, p‐NF‐κB p65 and NF‐κB p65 protein expressions ( n = 3). (E) BV2 cells were treated with LPS, Fc, ARTN‐Fc, BAY 11‐7082, or ARTN‐Fc combined with TNF‐α, followed by immunofluorescence staining for p‐NF‐κB p65. p‐NF‐κB p65 is shown in green, and nuclei were counterstained with DAPI (blue). Scale bar = 50 μm; magnified images, scale bar = 5 μm. (F) Representative Western blot images showing the protein levels of TLR4, p‐NF‐κB p65, and NF‐κB p65 in BV2 cells from each group. β‐actin was used as an endogenous control. (G, H) Quantitative analysis of TLR4 protein expression and the p‐NF‐κB p65/NF‐κB p65 ratio ( n = 3). Statistical analysis was performed using one‐way ANOVA followed by Tukey's multiple comparisons test. * p < 0.05, ** p < 0.01, n.s. = not significant.

Article Snippet: Lipopolysaccharide (HY‐D1056), TNF‐α (HY‐P1860) and BAY 11‐7082 (HY‐13453) were purchased from MedChemExpress (NJ, USA).

Techniques: Translocation Assay, Immunofluorescence, Fluorescence, Marker, Western Blot, Staining, Control, Expressing