ampk  (Proteintech)

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: Bioactive Materials

Article Title: A promising magnesium-related alloy with metabolic reprogramming and antitumor effects in hepatocellular and pancreatic cancer

doi: 10.1016/j.bioactmat.2025.12.039

Figure Lengend Snippet: Transcriptomic Analysis Indicates Mg and Al-Mg Alter Gene Expression Patterns in Hepatocellular and Pancreatic Cancer Cells. (A) High-throughput sequencing of PANC-1, PANC-1-Mg, PANC-1-Al-Mg, Huh7, Huh7-Mg, and Huh7-Al-Mg groups; heatmap showing sample correlations. (B) Volcano plots illustrating differential gene expression in Huh7 and PANC-1 cells after Mg or Al-Mg treatment. (C) GO enrichment analysis of differentially expressed genes following Mg or Al-Mg treatment. (D) KEGG pathway enrichment analysis of differentially expressed genes after Mg or Al-Mg treatment. (E) GSEA enrichment analysis of gene expression profiles post Mg or Al-Mg treatment. (F) Western blot analysis of AMPK, p-AMPK and CPT1B expression in PANC-1 cells after Mg or Al-Mg exposure with or without AMPK agonist. (G) PPI network analysis of differentially expressed genes in Mg or Al-Mg-treated cells. (H) Integrated analysis of metabolomic and transcriptomic data by MetaboAnalyst 6.0.

Article Snippet: After blocking with 5 % nonfat milk for 1 h at room temperature, membranes were incubated overnight at 4 °C with primary antibodies, including AMPK (1:1000, CST, 2532), p-AMPK (1:1000, CST, 2535), CPT1B (1:1000, Proteintech, 22170-1-AP), CDK4 (1:1000, Proteintech, 11026-1-AP), PCNA (1:1000, Proteintech, 10205-2-AP), p21 (1:1000, Proteintech, 10355-1-AP), GAPDH (1:1000, Proteintech, 60004-1-Ig) followed by HRP-conjugated secondary antibody (1:5000, Proteintech, RGAR001) for 1 h at room temperature.

Techniques: Gene Expression, Next-Generation Sequencing, Western Blot, Expressing

Journal: Bioactive Materials

Article Title: A promising magnesium-related alloy with metabolic reprogramming and antitumor effects in hepatocellular and pancreatic cancer

doi: 10.1016/j.bioactmat.2025.12.039

Figure Lengend Snippet: Transcriptomic Analysis Indicates Mg and Al-Mg Alter Gene Expression Patterns in Hepatocellular and Pancreatic Cancer Cells. (A) High-throughput sequencing of PANC-1, PANC-1-Mg, PANC-1-Al-Mg, Huh7, Huh7-Mg, and Huh7-Al-Mg groups; heatmap showing sample correlations. (B) Volcano plots illustrating differential gene expression in Huh7 and PANC-1 cells after Mg or Al-Mg treatment. (C) GO enrichment analysis of differentially expressed genes following Mg or Al-Mg treatment. (D) KEGG pathway enrichment analysis of differentially expressed genes after Mg or Al-Mg treatment. (E) GSEA enrichment analysis of gene expression profiles post Mg or Al-Mg treatment. (F) Western blot analysis of AMPK, p-AMPK and CPT1B expression in PANC-1 cells after Mg or Al-Mg exposure with or without AMPK agonist. (G) PPI network analysis of differentially expressed genes in Mg or Al-Mg-treated cells. (H) Integrated analysis of metabolomic and transcriptomic data by MetaboAnalyst 6.0.

Article Snippet: Primary antibodies used included p-AMPK (1:100, CST, 2535), Ki-67 (1:1000, Servicebio, GB111499-50), E-cadherin (1:500, Servicebio, GB12083-100), N-cadherin (1:500, Proteintech, SA00013-4), and Vimentin (1:2000, Servicebio, GB11192-100).

Techniques: Gene Expression, Next-Generation Sequencing, Western Blot, Expressing

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Western blot analysis of female mouse livers fed HFD containing doxycycline (Doxy) to express constitutive active AMPK [a1(1-312); AMPK CA ] for 7 days compared to control livers. ( B ) Immunohistochemical analysis of livers from male control and AMPK CA mice fed HFD+Doxy for 7 months. Scale bars, 100 μm. Right: Quantification of the staining intensity for P-Thr 172 AMPK and P-Ser 79 ACC staining. n = 6 to 10. ±SEM, Welch t test. ( C ) Representative whole-mount (top) and MRI (bottom) image of male control and AMPK CA livers 9 months post–DEN injection. Yellow dashed lines indicate tumors. ( D ) Quantification of the tumor number in male control and AMPK CA livers 6 and 9 months post–DEN injection based on MRI images. 6 months: N = 5 to 7; 9 months: N = 16 to 17; Fisher least significant difference (LSD) test. ( E ) Quantification of the serum AFP levels in control and AMPK CA male mice 6 and 9 months post–DEN injection. n = 9 to 15. Fisher LSD test. * P < 0.05; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Western Blot, Control, Immunohistochemical staining, Staining, Injection

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Quantification of the tumor size from MRI imaging at 6 and 9 months. n = 10 to 448. Log normal Welch t test. ( B ) Immunohistochemical analysis of BrdU incorporation in tumor tissue from control and AMPK CA livers following a 4-hour BrdU pulse (10 mg/kg). Scale bars, 100 μm. Right: Quantification of the number of positive cells per area. n = 13 to 17. Welch t test. ( C ) Gene expression of Ampka1 normalized to Actin in nontumor and tumor tissue. n = 5 to 6. ±SEM, Fisher LSD test. ( D ) Immunohistochemical analysis of P-Ser 79 ACC in nontumor and tumor tissue from control and AMPK CA livers. Scale bars, 100 μm. ( E ) Quantification of the staining intensity of P-Ser 79 ACC. n = 5 to 14. Fisher LSD test. ( F ) Immunohistochemical analysis of P-Thr 172 AMPK in nontumor and tumor tissue from control and AMPK CA livers. Scale bars, 100 μm. ( G ) Quantification of the staining intensity. n = 6 to 11. Fisher LSD test. ( H ) Western blot analysis of nontumor (NT) and tumor (T) tissue. VINCULIN as loading control. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Imaging, Immunohistochemical staining, BrdU Incorporation Assay, Control, Gene Expression, Staining, Western Blot

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Weight of male control and AMPK CA mice on HFD for 8 and 16 weeks following STZ injection n = 14 to 19. Fisher LSD test. ( B ) Fasting blood glucose levels 20 weeks post–STZ injection. n = 6 to 8. Welch t test. ( C ) Oil Red O staining of control and AMPK CA livers 20 weeks following STZ injection. Right: Scoring of Oil Red O levels. n = 7. Welch t test. Scale bar, 100 μm. ( D ) Quantification of the triglyceride levels in livers of control and AMPK CA mice 20 weeks post–STZ injection. n = 5. Welch t test. ( E ) Whole-mount image (top) and MRI image (bottom) of livers 20 weeks following STZ injection. Yellow dashed lines indicate tumors. ( F ) Quantification of the tumor number from control and AMPK CA mice injected with STZ. n = 8. Welch t test. ( G ) Liver weight relative to body weight of STZ-injected mice. n = 13 to 14. Welch t test. ( H ) Quantification of tumor sizes 20 weeks following STZ injection. n = 67 to 136 from 13 to 14 mice. Log normal Welch t test. ( I ) Gene expression analysis of nontumor and tumor tissue from STZ-injected mice. n = 6 to 7. Fisher LSD test. ( J ) Western blot analysis of matched nontumor and tumor tissue from AMPK CA mice injected with STZ. Right: Quantification of total and phospho-AMPK CA levels. n = 10 to 11. Welsh t test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Control, Injection, Staining, Gene Expression, Western Blot

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Quantification of the relative levels of bile acids in livers from control and AMPK CA mice following 2 months (2 m) of doxycycline. n = 5. Welsh t test. ( B ) Heatmap of average relative bile acids levels of AMPK CA livers relative to control livers following varying durations of doxycycline treatment [15 days (15d) and 2 months] or in matched nontumor and tumor tissues [7 months (7 m) and “Tumor”], n = 4 to 6 per genotype per time point. ( C ) Ratio of conjugated versus nonconjugated bile acids in livers from control or AMPK CA mice on chow or HFD for 15 days. n = 4 to 6. Welsh t test. ( D ) Ratio of secondary (DCA;LCA) versus primary (CA;CDCA) bile acids in livers from control or AMPK CA mice on chow or HFD for 15 days. n = 4 to 6. Fisher LSD test. DCA, deoxycholic acid; LCA, lithocholic acid; CA, cholic acid; CDCA, chenodeoxycholic acid. ( E ) Quantification of the serum bile acid levels relative to control livers of tumor bearing mice on doxycycline diet for 7 months. n = 5 to 6. Welch t test. ( F ) Quantification of the TBARS concentration in livers from control or AMPK CA mice on chow or HFD containing doxycycline for 15 days or 2 months. n = 4 to 6. Fisher LSD test. ( G ) Quantification of the gene expression in control or AMPK CA livers of mice on chow or HFD containing doxycycline for 3 or 15 days or in tumors. n = 4 to 6. Fisher LSD test. ( H ) Western blot analysis of livers from mice on HFD containing doxycycline for 15 days. ( I ) Gene expression of YAP target genes in WT primary hepatocytes treated with 80 μM TLCA or DMSO for 6 hours. n = 5 to 6. Two-way analysis of variance (ANOVA). ( J ) Western blot analysis of primary hepatocytes treated with 80 μM TLCA or DMSO for 30 min, 1 hour, or 2 hours. VINCULIN as loading control. n.s., not significant.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Control, Concentration Assay, Gene Expression, Western Blot

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Gene expression of bile acid biosynthesis proteins and bile acid response genes in livers from mice fed HFD containing doxycycline for 15 days. n = 4. Fisher LSD test. ( B ) Heatmap of the log 2 fold change of gene expression changes in AMPK CA relative to control livers from mice on HFD containing doxycycline for 2 months. ( C ) Gene expression of bile acid transporters in livers from mice fed chow or HFD containing doxycycline for 15 days. n = 4 to 6. Fisher LSD test. ( D ) Gene expression of bile acid transcription factor in livers from mice fed chow or HFD containing doxycycline for 15 days. n = 4 to 6. Fisher LSD test. ( E ) Gene expression of bile acid responsive genes in the ileum from mice fed HFD containing doxycycline for 15 days. n = 4. Fisher LSD test. ±SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Gene Expression, Control

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Transcription factors (TFs) enriched in AMPK CA mice relative to control mice following 2 months of HFD containing doxycycline using Enrichr and a 1.3-fold change with an adjusted P value less than 0.05. ( B ) Gene set enrichment analysis of genes in AMPK CA livers compared to control livers following 2 months of HFD containing doxycycline. NES, normalized enrichment score. ( C ) Western blot analysis of livers from control and AMPK CA mice following 15 days of HFD containing doxycycline. ( D ) Volcano plot of HNF4α binding sites altered in livers from control and AMPK CA mice following 15 days of HFD containing doxycycline. ( E ) Genomic location and directionality of change in HNF4α binding sites in livers from control and AMPK CA mice following 15 days of HFD containing doxycycline. TTS, transcriptional termination site. ( F ) Representative HNF4α binding sites in control versus AMPK CA livers. ( G ) Heatmap analysis of HNF4α target genes significantly changed (FDR < 0.05) in AMPK CA livers compared to control livers following 2 months of HFD containing doxycycline. Log 2 fold change. ( H ) Growth curve of the xenograft of MHCC-97H cells expressing WT or S304A or S304D mutant HNF4α. Inset: Representative image of tumor sizes at endpoint n = 11. ( I ) Weight of xenografts of MHCC-97H cells expressing WT or mutant HNF4α. n = 11. Welch t test. ( J ) Gene expression analysis of HNF4α targets in xenograft of MHCC-97H cells expressing WT or S304D mutant HNF4α. n = 11. ±SEM, Fisher LSD test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Control, Western Blot, Binding Assay, Expressing, Mutagenesis, Gene Expression