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amp activated protein kinase α ampkα  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc amp activated protein kinase α ampkα
    Amp Activated Protein Kinase α Ampkα, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 4497 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/ampk%CE%B1/AMPKalpha+Antibody/pmc12811471-130-16-22
    Average 97 stars, based on 4497 article reviews
    amp activated protein kinase α ampkα - by Bioz Stars, 2026-10
    97/100 stars

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    other:

    Article Title: AMPK inhibition and elevated angiogenin are associated with tRNA fragmentation in the male germline exposed to a high-fat diet
    Article Snippet: , AMPKα , Cell signaling technology , 2532 , 1:1000 in BSA.

    Article Title: Neuronal PPP2R5C in plasma is a potential biomarker for early diagnosis of Alzheimer’s disease
    Article Snippet: AMPKα , Cell Signaling Technology, #2603 , RRID: AB_490795.

    Expressing:

    Article Title: Enhancing autophagy or increasing longevity by administration of urolithins
    Article Snippet: .. Protein expression levels were examined for autophagy-related proteins LC3-I and LC3-II (Cell Signaling antibody #4108), p62 (Cell Signaling antibody #5114), AMPKα (Cell Signaling antibody #2603), and p-AMPKα (Cell Signaling antibody #2531). .. The housekeeping protein, B-actin (Cell Signaling Antibody #4967) level was measured as a loading control.

    Article Title: Enhancing autophagy or increasing longevity by administration of urolithins
    Article Snippet: .. Protein expression levels were examined for autophagy related proteins LC3-I and LC3-II (Cell Signaling antibody #4108), p62 (Cell Signalling antibody #5114), AMPKα (Cell Signaling antibody #2603), and p-AMPKα (Cell Signaling antibody #2531). ..

    Membrane:

    Article Title: Metformin drives HIF-1α–mediated dual metabolic reprogramming to enhance γδ T cell therapy in triple-negative breast cancer
    Article Snippet: Samples were resolved on 10% SDS–polyacrylamide gels and electrotransferred to 0.45-μm nitrocellulose membranes (IPVH00010, Merck millipore). .. The membrane was blocked with 5% non-fat milk in TBST (JXF003, Jingxin Bio) for 1 h at room temperature and subsequently incubated overnight at 4 °C with primary antibodies targeting phospho-AMPKα (2535 T, CST), AMPKα (2793S, CST), HIF-1α (2178S, CST), and β-actin (4970S, CST). .. The membrane was then subjected to three consecutive washes with TBST for 5 min, incubated for 1 h at room temperature with Anti-rabbit IgG HRP-linked Antibody (7074S, CST) or goat anti-mouse IgG HRP (abs20039, absin), washed three times in TBST for 5 min, and detected using the Omni-ECLTM Femto Light Chemiluminescence Kit (SQ201, EpiZyme).

    Article Title: Influence of diet-induced obesity and voluntary exercise training on cardiac lipids and mitochondrial function in mice
    Article Snippet: Densitometry was analyzed using ImageLab software (V6.0.1; Bio-Rad). .. Immunoblotting was performed with antibodies against lysocardiolipin acyltransferase 1(LCLAT1, PA5-25627; Thermo Fisher Scientific), PGC-1α (ab191838; Abcam, Cambridge, UK), MFN2 (PA5-118059; Thermo Fisher Scientific), ANP (sc-18811; Santa Cruz, Dallas, TX, USA), tumor necrosis factor alpha (TNFα, 3707; Cell Signaling, Danvers, MA, USA), phosphorylated adenosine monophosphate-activated protein kinase (AMPK, 2531; Cell Signaling), AMPKα (2532; Cell Signaling), Perilipin 5 (PA1-46215; Thermo Fisher Scientific), translocase of outer mitochondria membrane 70 (TOM70, 65675; Cell Signaling), cytochrome c oxidase subunit 4 (COXIV, 4844; Cell Signaling), and calnexin (208880; Merck, Rahway, NJ, USA). ..

    Article Title: Targeting GHSR-1α with silibinin to enhance mitophagy and prevent tubular injury in diabetic kidney disease.
    Article Snippet: Diabetic kidney disease (DKD) remains a leading cause of global renal failure, with tubular injury driven by mitochondrial dysfunction representing a critical yet therapeutically unaddressed mechanism.. Although the flavonolignan silibinin has demonstrated renoprotective potential, its precise molecular targets and mechanisms in DKD have remained unclear.. In this study, we combined integrated computational docking, biophysical assays, and comprehensive in vivo and in vitro models to investigate silibinin's interaction with the growth hormone secretagogue receptor-1α (GHSR-1α).

    Incubation:

    Article Title: Metformin drives HIF-1α–mediated dual metabolic reprogramming to enhance γδ T cell therapy in triple-negative breast cancer
    Article Snippet: Samples were resolved on 10% SDS–polyacrylamide gels and electrotransferred to 0.45-μm nitrocellulose membranes (IPVH00010, Merck millipore). .. The membrane was blocked with 5% non-fat milk in TBST (JXF003, Jingxin Bio) for 1 h at room temperature and subsequently incubated overnight at 4 °C with primary antibodies targeting phospho-AMPKα (2535 T, CST), AMPKα (2793S, CST), HIF-1α (2178S, CST), and β-actin (4970S, CST). .. The membrane was then subjected to three consecutive washes with TBST for 5 min, incubated for 1 h at room temperature with Anti-rabbit IgG HRP-linked Antibody (7074S, CST) or goat anti-mouse IgG HRP (abs20039, absin), washed three times in TBST for 5 min, and detected using the Omni-ECLTM Femto Light Chemiluminescence Kit (SQ201, EpiZyme).

    Western Blot:

    Article Title: Influence of diet-induced obesity and voluntary exercise training on cardiac lipids and mitochondrial function in mice
    Article Snippet: Densitometry was analyzed using ImageLab software (V6.0.1; Bio-Rad). .. Immunoblotting was performed with antibodies against lysocardiolipin acyltransferase 1(LCLAT1, PA5-25627; Thermo Fisher Scientific), PGC-1α (ab191838; Abcam, Cambridge, UK), MFN2 (PA5-118059; Thermo Fisher Scientific), ANP (sc-18811; Santa Cruz, Dallas, TX, USA), tumor necrosis factor alpha (TNFα, 3707; Cell Signaling, Danvers, MA, USA), phosphorylated adenosine monophosphate-activated protein kinase (AMPK, 2531; Cell Signaling), AMPKα (2532; Cell Signaling), Perilipin 5 (PA1-46215; Thermo Fisher Scientific), translocase of outer mitochondria membrane 70 (TOM70, 65675; Cell Signaling), cytochrome c oxidase subunit 4 (COXIV, 4844; Cell Signaling), and calnexin (208880; Merck, Rahway, NJ, USA). ..

    Pyrolysis Gas Chromatography:

    Article Title: Influence of diet-induced obesity and voluntary exercise training on cardiac lipids and mitochondrial function in mice
    Article Snippet: Densitometry was analyzed using ImageLab software (V6.0.1; Bio-Rad). .. Immunoblotting was performed with antibodies against lysocardiolipin acyltransferase 1(LCLAT1, PA5-25627; Thermo Fisher Scientific), PGC-1α (ab191838; Abcam, Cambridge, UK), MFN2 (PA5-118059; Thermo Fisher Scientific), ANP (sc-18811; Santa Cruz, Dallas, TX, USA), tumor necrosis factor alpha (TNFα, 3707; Cell Signaling, Danvers, MA, USA), phosphorylated adenosine monophosphate-activated protein kinase (AMPK, 2531; Cell Signaling), AMPKα (2532; Cell Signaling), Perilipin 5 (PA1-46215; Thermo Fisher Scientific), translocase of outer mitochondria membrane 70 (TOM70, 65675; Cell Signaling), cytochrome c oxidase subunit 4 (COXIV, 4844; Cell Signaling), and calnexin (208880; Merck, Rahway, NJ, USA). ..

    Aqueous Normal-phase Chromatography:

    Article Title: Influence of diet-induced obesity and voluntary exercise training on cardiac lipids and mitochondrial function in mice
    Article Snippet: Densitometry was analyzed using ImageLab software (V6.0.1; Bio-Rad). .. Immunoblotting was performed with antibodies against lysocardiolipin acyltransferase 1(LCLAT1, PA5-25627; Thermo Fisher Scientific), PGC-1α (ab191838; Abcam, Cambridge, UK), MFN2 (PA5-118059; Thermo Fisher Scientific), ANP (sc-18811; Santa Cruz, Dallas, TX, USA), tumor necrosis factor alpha (TNFα, 3707; Cell Signaling, Danvers, MA, USA), phosphorylated adenosine monophosphate-activated protein kinase (AMPK, 2531; Cell Signaling), AMPKα (2532; Cell Signaling), Perilipin 5 (PA1-46215; Thermo Fisher Scientific), translocase of outer mitochondria membrane 70 (TOM70, 65675; Cell Signaling), cytochrome c oxidase subunit 4 (COXIV, 4844; Cell Signaling), and calnexin (208880; Merck, Rahway, NJ, USA). ..

    Bioprocessing:

    Article Title: Dysregulation of a novel autophagosome-mitochondria contact contributes to autophagy dysfunction and neurodegeneration in tauopathy
    Article Snippet: .. Sources of antibodies or reagents are as follows: polyclonal antibodies against LC3 (Cell Signaling Technology, Cat# 2775), AMPKα (Cell Signaling Technology, Cat# 2532), p62/SQSTM1 (Abnova, Cat# H00008878-M01), NeuN (Millipore/Sigma, Cat# ABN78), TBC1D15 (Abcam, Cat# ab121396), Tau5 (DAKO, Cat# A0024), and synaptophysin/SYP (Abcam, Cat# ab32127); monoclonal antibodies against phospho-AMPKα (Cell Signaling Technology, Cat# 2535), AT8 (ThermoFisher Scientific, Cat# MN1020), syntaxin 1/STX1 (Santa Cruz Biotechnology, Cat# sc-12736), Rab7 (Sigma, Cat# R8779), GAPDH (Sigma, Cat# CB1001), TOM20 (Abcam, Cat# ab186734), mCherry (Takara, Cat# 632543), and Alexa fluor 488- (Cat# A-11017; Cat# A-11070), 546- (Cat# A-11018; Cat# A-11071), and 647- (Cat# A-21237; Cat# A-21246) conjugated secondary antibodies (Invitrogen); 10-NCP (VWR, Cat# 80017-188), trehalose (Cat# T0167), cycloheximide (Cat# 01810), rotenone (Cat# 557368), antimycin A (Cat# 1397-94-0), AICAR (Cat# A9978), CC (Cat# 171264), epoxomicin (Cat# 324800), CID 1067700 (Cat# SML0545), DMSO (Cat# D2650), and Sarkosyl (Cat# L9150) (Sigma); TBC1D15 shRNA (Cat# sc-154093-SH), Fis1 shRNA (Cat# sc-60644-SH), and control shRNA (Cat# sc-108060) (Santa Cruz Biotechnology); MitoView TM Green (Cat# 70054) (biotium); LentiBrite RFP-LC3 Lentiviral Biosensor (Cat# 17-10143) (Sigma). .. The constructs encoding GFP-LC3, DsRed-Mito, and mRFP-LC3 were prepared as we previously described ( , , , ). mCherry-TBC1D15 and mCherry-TBC1D15 D397A were from Vector Biolabs.

    shRNA:

    Article Title: Dysregulation of a novel autophagosome-mitochondria contact contributes to autophagy dysfunction and neurodegeneration in tauopathy
    Article Snippet: .. Sources of antibodies or reagents are as follows: polyclonal antibodies against LC3 (Cell Signaling Technology, Cat# 2775), AMPKα (Cell Signaling Technology, Cat# 2532), p62/SQSTM1 (Abnova, Cat# H00008878-M01), NeuN (Millipore/Sigma, Cat# ABN78), TBC1D15 (Abcam, Cat# ab121396), Tau5 (DAKO, Cat# A0024), and synaptophysin/SYP (Abcam, Cat# ab32127); monoclonal antibodies against phospho-AMPKα (Cell Signaling Technology, Cat# 2535), AT8 (ThermoFisher Scientific, Cat# MN1020), syntaxin 1/STX1 (Santa Cruz Biotechnology, Cat# sc-12736), Rab7 (Sigma, Cat# R8779), GAPDH (Sigma, Cat# CB1001), TOM20 (Abcam, Cat# ab186734), mCherry (Takara, Cat# 632543), and Alexa fluor 488- (Cat# A-11017; Cat# A-11070), 546- (Cat# A-11018; Cat# A-11071), and 647- (Cat# A-21237; Cat# A-21246) conjugated secondary antibodies (Invitrogen); 10-NCP (VWR, Cat# 80017-188), trehalose (Cat# T0167), cycloheximide (Cat# 01810), rotenone (Cat# 557368), antimycin A (Cat# 1397-94-0), AICAR (Cat# A9978), CC (Cat# 171264), epoxomicin (Cat# 324800), CID 1067700 (Cat# SML0545), DMSO (Cat# D2650), and Sarkosyl (Cat# L9150) (Sigma); TBC1D15 shRNA (Cat# sc-154093-SH), Fis1 shRNA (Cat# sc-60644-SH), and control shRNA (Cat# sc-108060) (Santa Cruz Biotechnology); MitoView TM Green (Cat# 70054) (biotium); LentiBrite RFP-LC3 Lentiviral Biosensor (Cat# 17-10143) (Sigma). .. The constructs encoding GFP-LC3, DsRed-Mito, and mRFP-LC3 were prepared as we previously described ( , , , ). mCherry-TBC1D15 and mCherry-TBC1D15 D397A were from Vector Biolabs.

    Control:

    Article Title: Dysregulation of a novel autophagosome-mitochondria contact contributes to autophagy dysfunction and neurodegeneration in tauopathy
    Article Snippet: .. Sources of antibodies or reagents are as follows: polyclonal antibodies against LC3 (Cell Signaling Technology, Cat# 2775), AMPKα (Cell Signaling Technology, Cat# 2532), p62/SQSTM1 (Abnova, Cat# H00008878-M01), NeuN (Millipore/Sigma, Cat# ABN78), TBC1D15 (Abcam, Cat# ab121396), Tau5 (DAKO, Cat# A0024), and synaptophysin/SYP (Abcam, Cat# ab32127); monoclonal antibodies against phospho-AMPKα (Cell Signaling Technology, Cat# 2535), AT8 (ThermoFisher Scientific, Cat# MN1020), syntaxin 1/STX1 (Santa Cruz Biotechnology, Cat# sc-12736), Rab7 (Sigma, Cat# R8779), GAPDH (Sigma, Cat# CB1001), TOM20 (Abcam, Cat# ab186734), mCherry (Takara, Cat# 632543), and Alexa fluor 488- (Cat# A-11017; Cat# A-11070), 546- (Cat# A-11018; Cat# A-11071), and 647- (Cat# A-21237; Cat# A-21246) conjugated secondary antibodies (Invitrogen); 10-NCP (VWR, Cat# 80017-188), trehalose (Cat# T0167), cycloheximide (Cat# 01810), rotenone (Cat# 557368), antimycin A (Cat# 1397-94-0), AICAR (Cat# A9978), CC (Cat# 171264), epoxomicin (Cat# 324800), CID 1067700 (Cat# SML0545), DMSO (Cat# D2650), and Sarkosyl (Cat# L9150) (Sigma); TBC1D15 shRNA (Cat# sc-154093-SH), Fis1 shRNA (Cat# sc-60644-SH), and control shRNA (Cat# sc-108060) (Santa Cruz Biotechnology); MitoView TM Green (Cat# 70054) (biotium); LentiBrite RFP-LC3 Lentiviral Biosensor (Cat# 17-10143) (Sigma). .. The constructs encoding GFP-LC3, DsRed-Mito, and mRFP-LC3 were prepared as we previously described ( , , , ). mCherry-TBC1D15 and mCherry-TBC1D15 D397A were from Vector Biolabs.



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    Taurine is the key small molecule in A1TP-HX-EVs that activated the <t>AMPK/NRF2</t> pathway to regulate nucleus pulposus cell repair. (A) The LC-MS/MS analysis was used to detect the differential active small molecule components between placental HX-EVs and EVs. (B) The SMPDB enrichment analysis identified pathways related to small molecules that are up-expressed in HX-EVs compared to EVs. The metabolic pathways marked in red are related to ferroptosis inhibition and mitochondrial function. (C) Volcano plot of small molecule in HX-EVs versus EVs. |log2FC| > 0.5, FDR <0.05. (D) The content of taurine in placental MSC (pMSC), hypoxia-induced pMSC(HX-pMSC) and their derived EVs was detected by ELISA. n = 3. (E) Primary NPCs cells were induced with TBHP, and then treated with EVs, HX-EVs, and A1TP-HX-EVs for 24 h. The cell lysates were subjected to ELISA assay to detect taurine content. (F) Two shRNA lentiviruses were designed to knock down TAUT a key enzyme in taurine uptake in pMSC. (G) The content of taurine in TAUT-sh1-pMSC and TAUT-sh2-pMSC derived EVs (KD-HX-EVs) was detected by ELISA. n = 3. (H) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (I) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h, followed by immunofluorescent staining with anti- TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (J) <t>A</t> <t>CDO1-overexpressing</t> retrovirus was designed to overexpress CDO1 in pMSCs. (K) The content of taurine in CDO1-OE-pMSC derived EVs (OE-EVs) was detected by ELISA. n = 3. (L) Primary NPCs were induced with TBHP, and then treated with treated A1TP-EVs and A1TP-OE-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (M) Primary NPCs were induced with TBHP, and then treated with A1TP-EVs and A1TP-OE-EVs for 24 h, followed by immunofluorescent staining with anti-TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (N-O) Representative oxygen consumption traces of primary NPCs induced with TBHP and then treated with A1TP-HX-EVs, A1TP-KD-HX-EVs, or A1TP-OE-EVs for 24 h. Maximal respiration of NPCs were quantified. n = 3. All data are expressed as the mean ± SD. For E), I), M) and O), one‐way ANOVA with Tukey's multiple comparison tests were used for statistical analysis. For D), G) and K), two‐tailed unpaired Student's t‐tests were used for statistical analysis. ∗ P < 0.05. ∗∗ P < 0.01. ∗∗∗ P < 0.001. ns, not significant.
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    Taurine is the key small molecule in A1TP-HX-EVs that activated the <t>AMPK/NRF2</t> pathway to regulate nucleus pulposus cell repair. (A) The LC-MS/MS analysis was used to detect the differential active small molecule components between placental HX-EVs and EVs. (B) The SMPDB enrichment analysis identified pathways related to small molecules that are up-expressed in HX-EVs compared to EVs. The metabolic pathways marked in red are related to ferroptosis inhibition and mitochondrial function. (C) Volcano plot of small molecule in HX-EVs versus EVs. |log2FC| > 0.5, FDR <0.05. (D) The content of taurine in placental MSC (pMSC), hypoxia-induced pMSC(HX-pMSC) and their derived EVs was detected by ELISA. n = 3. (E) Primary NPCs cells were induced with TBHP, and then treated with EVs, HX-EVs, and A1TP-HX-EVs for 24 h. The cell lysates were subjected to ELISA assay to detect taurine content. (F) Two shRNA lentiviruses were designed to knock down TAUT a key enzyme in taurine uptake in pMSC. (G) The content of taurine in TAUT-sh1-pMSC and TAUT-sh2-pMSC derived EVs (KD-HX-EVs) was detected by ELISA. n = 3. (H) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (I) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h, followed by immunofluorescent staining with anti- TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (J) <t>A</t> <t>CDO1-overexpressing</t> retrovirus was designed to overexpress CDO1 in pMSCs. (K) The content of taurine in CDO1-OE-pMSC derived EVs (OE-EVs) was detected by ELISA. n = 3. (L) Primary NPCs were induced with TBHP, and then treated with treated A1TP-EVs and A1TP-OE-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (M) Primary NPCs were induced with TBHP, and then treated with A1TP-EVs and A1TP-OE-EVs for 24 h, followed by immunofluorescent staining with anti-TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (N-O) Representative oxygen consumption traces of primary NPCs induced with TBHP and then treated with A1TP-HX-EVs, A1TP-KD-HX-EVs, or A1TP-OE-EVs for 24 h. Maximal respiration of NPCs were quantified. n = 3. All data are expressed as the mean ± SD. For E), I), M) and O), one‐way ANOVA with Tukey's multiple comparison tests were used for statistical analysis. For D), G) and K), two‐tailed unpaired Student's t‐tests were used for statistical analysis. ∗ P < 0.05. ∗∗ P < 0.01. ∗∗∗ P < 0.001. ns, not significant.
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    Taurine is the key small molecule in A1TP-HX-EVs that activated the <t>AMPK/NRF2</t> pathway to regulate nucleus pulposus cell repair. (A) The LC-MS/MS analysis was used to detect the differential active small molecule components between placental HX-EVs and EVs. (B) The SMPDB enrichment analysis identified pathways related to small molecules that are up-expressed in HX-EVs compared to EVs. The metabolic pathways marked in red are related to ferroptosis inhibition and mitochondrial function. (C) Volcano plot of small molecule in HX-EVs versus EVs. |log2FC| > 0.5, FDR <0.05. (D) The content of taurine in placental MSC (pMSC), hypoxia-induced pMSC(HX-pMSC) and their derived EVs was detected by ELISA. n = 3. (E) Primary NPCs cells were induced with TBHP, and then treated with EVs, HX-EVs, and A1TP-HX-EVs for 24 h. The cell lysates were subjected to ELISA assay to detect taurine content. (F) Two shRNA lentiviruses were designed to knock down TAUT a key enzyme in taurine uptake in pMSC. (G) The content of taurine in TAUT-sh1-pMSC and TAUT-sh2-pMSC derived EVs (KD-HX-EVs) was detected by ELISA. n = 3. (H) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (I) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h, followed by immunofluorescent staining with anti- TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (J) <t>A</t> <t>CDO1-overexpressing</t> retrovirus was designed to overexpress CDO1 in pMSCs. (K) The content of taurine in CDO1-OE-pMSC derived EVs (OE-EVs) was detected by ELISA. n = 3. (L) Primary NPCs were induced with TBHP, and then treated with treated A1TP-EVs and A1TP-OE-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (M) Primary NPCs were induced with TBHP, and then treated with A1TP-EVs and A1TP-OE-EVs for 24 h, followed by immunofluorescent staining with anti-TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (N-O) Representative oxygen consumption traces of primary NPCs induced with TBHP and then treated with A1TP-HX-EVs, A1TP-KD-HX-EVs, or A1TP-OE-EVs for 24 h. Maximal respiration of NPCs were quantified. n = 3. All data are expressed as the mean ± SD. For E), I), M) and O), one‐way ANOVA with Tukey's multiple comparison tests were used for statistical analysis. For D), G) and K), two‐tailed unpaired Student's t‐tests were used for statistical analysis. ∗ P < 0.05. ∗∗ P < 0.01. ∗∗∗ P < 0.001. ns, not significant.
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    Taurine is the key small molecule in A1TP-HX-EVs that activated the <t>AMPK/NRF2</t> pathway to regulate nucleus pulposus cell repair. (A) The LC-MS/MS analysis was used to detect the differential active small molecule components between placental HX-EVs and EVs. (B) The SMPDB enrichment analysis identified pathways related to small molecules that are up-expressed in HX-EVs compared to EVs. The metabolic pathways marked in red are related to ferroptosis inhibition and mitochondrial function. (C) Volcano plot of small molecule in HX-EVs versus EVs. |log2FC| > 0.5, FDR <0.05. (D) The content of taurine in placental MSC (pMSC), hypoxia-induced pMSC(HX-pMSC) and their derived EVs was detected by ELISA. n = 3. (E) Primary NPCs cells were induced with TBHP, and then treated with EVs, HX-EVs, and A1TP-HX-EVs for 24 h. The cell lysates were subjected to ELISA assay to detect taurine content. (F) Two shRNA lentiviruses were designed to knock down TAUT a key enzyme in taurine uptake in pMSC. (G) The content of taurine in TAUT-sh1-pMSC and TAUT-sh2-pMSC derived EVs (KD-HX-EVs) was detected by ELISA. n = 3. (H) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (I) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h, followed by immunofluorescent staining with anti- TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (J) <t>A</t> <t>CDO1-overexpressing</t> retrovirus was designed to overexpress CDO1 in pMSCs. (K) The content of taurine in CDO1-OE-pMSC derived EVs (OE-EVs) was detected by ELISA. n = 3. (L) Primary NPCs were induced with TBHP, and then treated with treated A1TP-EVs and A1TP-OE-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (M) Primary NPCs were induced with TBHP, and then treated with A1TP-EVs and A1TP-OE-EVs for 24 h, followed by immunofluorescent staining with anti-TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (N-O) Representative oxygen consumption traces of primary NPCs induced with TBHP and then treated with A1TP-HX-EVs, A1TP-KD-HX-EVs, or A1TP-OE-EVs for 24 h. Maximal respiration of NPCs were quantified. n = 3. All data are expressed as the mean ± SD. For E), I), M) and O), one‐way ANOVA with Tukey's multiple comparison tests were used for statistical analysis. For D), G) and K), two‐tailed unpaired Student's t‐tests were used for statistical analysis. ∗ P < 0.05. ∗∗ P < 0.01. ∗∗∗ P < 0.001. ns, not significant.
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    Taurine is the key small molecule in A1TP-HX-EVs that activated the AMPK/NRF2 pathway to regulate nucleus pulposus cell repair. (A) The LC-MS/MS analysis was used to detect the differential active small molecule components between placental HX-EVs and EVs. (B) The SMPDB enrichment analysis identified pathways related to small molecules that are up-expressed in HX-EVs compared to EVs. The metabolic pathways marked in red are related to ferroptosis inhibition and mitochondrial function. (C) Volcano plot of small molecule in HX-EVs versus EVs. |log2FC| > 0.5, FDR <0.05. (D) The content of taurine in placental MSC (pMSC), hypoxia-induced pMSC(HX-pMSC) and their derived EVs was detected by ELISA. n = 3. (E) Primary NPCs cells were induced with TBHP, and then treated with EVs, HX-EVs, and A1TP-HX-EVs for 24 h. The cell lysates were subjected to ELISA assay to detect taurine content. (F) Two shRNA lentiviruses were designed to knock down TAUT a key enzyme in taurine uptake in pMSC. (G) The content of taurine in TAUT-sh1-pMSC and TAUT-sh2-pMSC derived EVs (KD-HX-EVs) was detected by ELISA. n = 3. (H) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (I) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h, followed by immunofluorescent staining with anti- TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (J) A CDO1-overexpressing retrovirus was designed to overexpress CDO1 in pMSCs. (K) The content of taurine in CDO1-OE-pMSC derived EVs (OE-EVs) was detected by ELISA. n = 3. (L) Primary NPCs were induced with TBHP, and then treated with treated A1TP-EVs and A1TP-OE-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (M) Primary NPCs were induced with TBHP, and then treated with A1TP-EVs and A1TP-OE-EVs for 24 h, followed by immunofluorescent staining with anti-TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (N-O) Representative oxygen consumption traces of primary NPCs induced with TBHP and then treated with A1TP-HX-EVs, A1TP-KD-HX-EVs, or A1TP-OE-EVs for 24 h. Maximal respiration of NPCs were quantified. n = 3. All data are expressed as the mean ± SD. For E), I), M) and O), one‐way ANOVA with Tukey's multiple comparison tests were used for statistical analysis. For D), G) and K), two‐tailed unpaired Student's t‐tests were used for statistical analysis. ∗ P < 0.05. ∗∗ P < 0.01. ∗∗∗ P < 0.001. ns, not significant.

    Journal: Bioactive Materials

    Article Title: ADGRG1-targeted hypoxia preconditioned extracellular vesicles ameliorate intervertebral disc degeneration by delivering taurine to disrupt the oxidative stress feedback loop-driven ferroptosis in nucleus pulposus cells

    doi: 10.1016/j.bioactmat.2026.02.029

    Figure Lengend Snippet: Taurine is the key small molecule in A1TP-HX-EVs that activated the AMPK/NRF2 pathway to regulate nucleus pulposus cell repair. (A) The LC-MS/MS analysis was used to detect the differential active small molecule components between placental HX-EVs and EVs. (B) The SMPDB enrichment analysis identified pathways related to small molecules that are up-expressed in HX-EVs compared to EVs. The metabolic pathways marked in red are related to ferroptosis inhibition and mitochondrial function. (C) Volcano plot of small molecule in HX-EVs versus EVs. |log2FC| > 0.5, FDR <0.05. (D) The content of taurine in placental MSC (pMSC), hypoxia-induced pMSC(HX-pMSC) and their derived EVs was detected by ELISA. n = 3. (E) Primary NPCs cells were induced with TBHP, and then treated with EVs, HX-EVs, and A1TP-HX-EVs for 24 h. The cell lysates were subjected to ELISA assay to detect taurine content. (F) Two shRNA lentiviruses were designed to knock down TAUT a key enzyme in taurine uptake in pMSC. (G) The content of taurine in TAUT-sh1-pMSC and TAUT-sh2-pMSC derived EVs (KD-HX-EVs) was detected by ELISA. n = 3. (H) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (I) Primary NPCs were induced with TBHP, and then treated with A1TP-HX-EVs and A1TP-KD-HX-EVs for 24 h, followed by immunofluorescent staining with anti- TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (J) A CDO1-overexpressing retrovirus was designed to overexpress CDO1 in pMSCs. (K) The content of taurine in CDO1-OE-pMSC derived EVs (OE-EVs) was detected by ELISA. n = 3. (L) Primary NPCs were induced with TBHP, and then treated with treated A1TP-EVs and A1TP-OE-EVs for 24 h. Cell lysates were immunoblotted with indicated antibodies. (M) Primary NPCs were induced with TBHP, and then treated with A1TP-EVs and A1TP-OE-EVs for 24 h, followed by immunofluorescent staining with anti-TOM20 (green) and anti-4-HNE (red) antibodies. n = 3. Scale bar, 50 μm. (N-O) Representative oxygen consumption traces of primary NPCs induced with TBHP and then treated with A1TP-HX-EVs, A1TP-KD-HX-EVs, or A1TP-OE-EVs for 24 h. Maximal respiration of NPCs were quantified. n = 3. All data are expressed as the mean ± SD. For E), I), M) and O), one‐way ANOVA with Tukey's multiple comparison tests were used for statistical analysis. For D), G) and K), two‐tailed unpaired Student's t‐tests were used for statistical analysis. ∗ P < 0.05. ∗∗ P < 0.01. ∗∗∗ P < 0.001. ns, not significant.

    Article Snippet: Primary antibodies included FTH1(4393S, Cell Signaling Technology), COL1A1(72026T, Cell Signaling Technology), COL2A1(sc-52658, Santa Cruz Biotechnology), MMP13(ab39012, Abcam), GPX4(30388-1-AP, Proteintech), ADGRG1(sc-390192, Santa Cruz Biotechnology), TAUT (sc-393036, Santa Cruz Biotechnology), TonEBP (sc-101098, Santa Cruz Biotechnology), CDO1 (12589-1-AP, Proteintech), AMPK(10929-2-AP, Proteintech), Phospho-AMPK (Thr172)(2535T, Cell Signaling Technology), SIRT1(8469T, Cell Signaling Technology), P-SIRT1(Ser47)(2314S, Cell Signaling Technology), PGC-1α(2178S, Cell Signaling Technology), Ac-lysine(sc-81623, Santa Cruz Biotechnology), NRF2(16396-1-AP, Proteintech), TFAM(22586-1-AP, Proteintech), NCOA4(66849S, Santa Cruz Biotechnology).

    Techniques: Liquid Chromatography with Mass Spectroscopy, Inhibition, Derivative Assay, Enzyme-linked Immunosorbent Assay, shRNA, Knockdown, Staining, Comparison, Two Tailed Test