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nape pld  (Santa Cruz Biotechnology)


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    Structured Review

    Santa Cruz Biotechnology nape pld
    Nape Pld, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nape+pld/NAPE-PLD+Antibody/pm40706771-74-31-34
    Average 93 stars, based on 10 article reviews
    nape pld - by Bioz Stars, 2026-10
    93/100 stars

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    Related Articles

    Incubation:

    Article Title: Long-Term Tamoxifen Effects in the Cyclic Interaction of the Endocannabinoid and Endocrine System in the Rat Central Nervous System.
    Article Snippet: .. Subsequently, slides were incubated overnight at 4 °C, with the primary antibodies against CB1, CB2, NAPE-PLD and FAAH (Table 1), all from St. Cruz Biotechnology (Santa Cruz, CA, USA). .. After washing, they were incubated with diluted biotinylated secondary antibodies (Table 1) for 1 h, followed by incubation with avidin-biotin peroxidase complex (Vectastain Elite ABC Kit) from Vector Laboratories (Burlingame, CA, USA).

    Article Title: Long-Term Tamoxifen Effects in the Cyclic Interaction of the Endocannabinoid and Endocrine System in the Rat Central Nervous System.
    Article Snippet: .. Subsequently, slides were incubated overnight at 4 ◦C, with the primary antibodies against CB1, CB2, NAPE-PLD and FAAH (Table 1), all from St. Cruz Biotechnology (Santa Cruz, CA, USA). .. After washing, they were incubated with diluted biotinylated secondary antibodies (Table 1) for 1 h, followed by incubation with avidin-biotin peroxidase complex (Vectastain Elite ABC Kit) from Vector Laboratories (Burlingame, CA, USA).

    Article Title: Effects of cannabidiol, cannabichromene, cannabidivarin, cannabigerol and cannabinol in endometrial cells: Implications for endocrine and senescence modulation.
    Article Snippet: .. Membranes were incubated with mouse monoclonal antibodies against ER-α (1:200, sc-8002; Santa Cruz Biotechnology, CA, USA), AR (1:200, sc-7305; Santa Cruz Biotechnology, CA, USA), p-AR (1:400, ab45089; Abcam Limited, Cambridge, UK), NAPE-PLD (1:200, sc-514372; Santa Cruz Biotechnology, CA, USA) and FAAH (1:200, sc-100739; Santa Cruz Biotechnology, CA, USA) or rabbit monoclonal antibodies against p-p44/42 MAPK (p-ERK1/2) (Thr202/Tyr204) (1:200; 4370S, Cell Signaling Technology, WZ Leiden), p44/42 MAPK (ERK1/2) (1:200; 4695S, Cell Signaling Technology, WZ Leiden), p-AKT (Ser473) (1:100; 4060S, Cell Signaling Technology, WZ Leiden) and AKT (1:100; 4691S, Cell Signaling Technology, WZ Leiden), at 4 oC overnight. .. Then, membranes were washed and incubated with peroxidase-conjugated secondary antibody antimouse or anti-rabbit (1:2000 or 1:4000 in the case of p-AR) acquired from Thermo Fisher, Waltham, MA, USA.

    Article Title: Long-Term Tamoxifen Effects in the Cyclic Interaction of the Endocannabinoid and Endocrine System in the Rat Central Nervous System
    Article Snippet: .. Subsequently, slides were incubated overnight at 4 °C, with the primary antibodies against CB1, CB2, NAPE-PLD and FAAH , all from St. Cruz Biotechnology (Santa Cruz, CA, USA). .. After washing, they were incubated with diluted biotinylated secondary antibodies ( ) for 1 h, followed by incubation with avidin-biotin peroxidase complex (Vectastain Elite ABC Kit) from Vector Laboratories (Burlingame, CA, USA).

    Article Title: Augmentation of Endogenous 2-Arachidonoylglycerol Mitigates Autistic Behaviors of BTBR Mice.
    Article Snippet: The lipid-based endocannabinoid (eCB) system regulates a host of developmental, physiological, and pathological processes in the mammalian brain, and recent studies have suggested that dysfunction of eCB system may contribute to the neuropathology of autism spectrum disorder (ASD).. However, specific contributions to ASD-related developmental, cognitive, and behavioral phenotypes remain largely unexplored.. The current study was designed to investigate if enhancing eCB signaling by blocking 2-arachidonoylglycerol (2-AG) hydrolase can mitigate ASD-like behaviors in a mouse model, and if such effects are associated with suppression of inflammatory signaling, oxidative stress, or neuronal apoptosis.

    Article Title: Discovery of a NAPE-PLD inhibitor that modulates emotional behavior in mice
    Article Snippet: Primary antibodies against NAPE-PLD (Abcam, ab95397, 1:200, in TBST), FLAG-tagged proteins (Sigma Aldrich, F3165, 1:5,000 in 5% milk in TBST), α-tubulin (Genetex, GTX76511, 1:5,000 in 5% milk in TBST) or β-actin (Abcam, ab8226, 1:5,000 in 5% milk in TBST) were incubated 1 h at room temperature or overnight at 4 °C. .. Membranes were washed with TBST and incubated with secondary antibodies: for NAPE-PLD, goat-antirabbit-HRP (Santa Cruz, sc-2030, 1:2,000 in 5% milk in TBST); for FLAG-tagged proteins and β-actin, goat-antimouse-HRP (Santa Cruz, sc-2005, 1:5,000 in 5% milk in TBST); for α-tubulin, goat-antirat (Santa Cruz, sc-2032, 1:5,000 in 5% milk in TBST). ..

    Article Title: R-Flurbiprofen Reduces Neuropathic Pain in Rodents by Restoring Endogenous Cannabinoids
    Article Snippet: Proteins were separated by SDS-PAGE (30 μg/lane) and transferred onto nitrocellulose membranes (Amersham Pharmacia Biotech) by wet-blotting. .. The blots were incubated with primary antibodies directed against CB1 (1∶500, Cayman Europe), CB2 (1∶500 Cayman Europe), FAAH (1∶1000, SantaCruz Biotechnology), NAPE-PLD (1∶500, Novus), Iba-1 (1∶500; Wako, Neuss, Germany), phospho-p38 and total p38 (Cell Signaling), CD68 (Abcam), beta-actin (SantaCruz Biotechnology), Hsp90 (SantaCruz Biotechnology) or extracellular signal-regulated kinase 2 (ERK-2; 1∶10000; SantaCruz Biotechnology) according to standard procedures. ..

    Bioprocessing:

    Article Title: Effects of cannabidiol, cannabichromene, cannabidivarin, cannabigerol and cannabinol in endometrial cells: Implications for endocrine and senescence modulation.
    Article Snippet: .. Membranes were incubated with mouse monoclonal antibodies against ER-α (1:200, sc-8002; Santa Cruz Biotechnology, CA, USA), AR (1:200, sc-7305; Santa Cruz Biotechnology, CA, USA), p-AR (1:400, ab45089; Abcam Limited, Cambridge, UK), NAPE-PLD (1:200, sc-514372; Santa Cruz Biotechnology, CA, USA) and FAAH (1:200, sc-100739; Santa Cruz Biotechnology, CA, USA) or rabbit monoclonal antibodies against p-p44/42 MAPK (p-ERK1/2) (Thr202/Tyr204) (1:200; 4370S, Cell Signaling Technology, WZ Leiden), p44/42 MAPK (ERK1/2) (1:200; 4695S, Cell Signaling Technology, WZ Leiden), p-AKT (Ser473) (1:100; 4060S, Cell Signaling Technology, WZ Leiden) and AKT (1:100; 4691S, Cell Signaling Technology, WZ Leiden), at 4 oC overnight. .. Then, membranes were washed and incubated with peroxidase-conjugated secondary antibody antimouse or anti-rabbit (1:2000 or 1:4000 in the case of p-AR) acquired from Thermo Fisher, Waltham, MA, USA.

    other:

    Article Title: Discovery of a NAPE-PLD inhibitor that modulates emotional behavior in mice
    Article Snippet: Protein concentrations were determined using a Bradford assay (Bio-Rad).



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    OriGene nape pld human gene knockout kit
    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking <t>down</t> <t>NAPE-PLD</t> using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.
    Nape Pld Human Gene Knockout Kit, supplied by OriGene, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking <t>down</t> <t>NAPE-PLD</t> using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.
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    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking <t>down</t> <t>NAPE-PLD</t> using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.
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    nape pld  (Santa Cruz Biotechnology)
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    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking <t>down</t> <t>NAPE-PLD</t> using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.
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    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking <t>down</t> <t>NAPE-PLD</t> using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.
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    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking <t>down</t> <t>NAPE-PLD</t> using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.
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    nape- pld wb  (Thermo Fisher)
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    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking <t>down</t> <t>NAPE-PLD</t> using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.
    Nape Pld Wb, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pld-nape 10306 antibody  (Cayman Chemical)
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    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking <t>down</t> <t>NAPE-PLD</t> using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.
    Pld Nape 10306 Antibody, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mouse anti nape pld  (Santa Cruz Biotechnology)
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    Santa Cruz Biotechnology mouse anti nape pld
    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking <t>down</t> <t>NAPE-PLD</t> using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.
    Mouse Anti Nape Pld, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nape+pld/NAPE-PLD+Antibody/pm39151841-141-55-60
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    Image Search Results


    ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking down NAPE-PLD using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.

    Journal: bioRxiv

    Article Title: A NAPE–LRRK2 metabolic axis controls lysosomal homeostasis in Parkinson’s disease

    doi: 10.64898/2026.03.19.712902

    Figure Lengend Snippet: ( A ) Schematic representation of the experimental strategy. Stable SH-SY5Y cell lines were generated either by knocking down NAPE-PLD using CRISP-Cas9 approach or by overexpressing PLA2G4E, both manipulations led to NAPE accumulation at the cell membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-knockdown (KD; violet bars) and wild-type (WT; blue bars) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in KD and WT cells. Top, representative blot; bottom, densitometric quantification normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. Nuclei were counterstained with DAPI. ( E ) Total NAPE levels in KD and WT cells. ( F ) PLA2G4E mRNA levels in PLA2G4E-overexpressing (OE; pink bars) and WT (blue bars) cells. ( G ) Western blot analysis of PLA2G4E protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( H ) Immunofluorescence analysis of PLA2G4E (gray) with representative images ( H1, H2 ) and quantification. Nuclei were counterstained with DAPI. ( I ) Total NAPE levels in OE and WT cells. Error bars represent SEM. *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001 Student’s t -test with Welch’s correction. Scale bars, 10 μm.

    Article Snippet: The NAPE-PLD Human Gene Knockout Kit (CRISPR, Origene, KN409877) was used following manufacturer’s instructions.

    Techniques: Generated, Knockdown, Western Blot, Immunofluorescence

    ( A ) Immunofluorescence analysis of LRRK2 (gray) with representative images ( A1–A3 ) and quantification in wild-type (WT; A1 , blue), NAPE-PLD knockdown (KD; A2 , violet), and PLA2G4E-overexpressing (OE; A3 , pink) SH-SY5Y cells. Nuclei were counterstained with DAPI. ( B ) Quantification of the colocalization between LAMP1-positive structures and LRRK2, expressed as Manders’ overlap coefficient for the fraction of LAMP1 signal overlapping with LRRK2. ( C ) Western blot analysis of Rab8a and phosphorylated Rab8a (pT72-Rab8a) in KD and OE compared with WT cells. Top, representative blot; bottom, densitometric quantification normalized to actin. ( D ) Representative super-resolution immunofluorescence images of LAMP1 (magenta) and LRRK2 (green) in WT ( D1 ), NAPE-PLD KD ( D2 ), and PLA2G4E OE ( D3 ) cells. Nuclei were stained with DAPI (gray). Scale bar, 5 μm. ( E ) Fluorescence intensity quantification and representative images of the Magic Red–cathepsin B assay, indicating lysosomal degradative capacity per cell, in WT ( E1 ), NAPE-PLD KD ( E2 ), and PLA2G4E OE ( E3 ) cells. Representative images are displayed using the FIRE lookup table (LUT) to indicate fluorescence intensity. Scale bars, 10 μm. Mean values are indicated in the graphs; error bars represent SEM. *P < 0.05, ****P < 0.0001; one-way ANOVA with Tukey’s multiple-comparison test.

    Journal: bioRxiv

    Article Title: A NAPE–LRRK2 metabolic axis controls lysosomal homeostasis in Parkinson’s disease

    doi: 10.64898/2026.03.19.712902

    Figure Lengend Snippet: ( A ) Immunofluorescence analysis of LRRK2 (gray) with representative images ( A1–A3 ) and quantification in wild-type (WT; A1 , blue), NAPE-PLD knockdown (KD; A2 , violet), and PLA2G4E-overexpressing (OE; A3 , pink) SH-SY5Y cells. Nuclei were counterstained with DAPI. ( B ) Quantification of the colocalization between LAMP1-positive structures and LRRK2, expressed as Manders’ overlap coefficient for the fraction of LAMP1 signal overlapping with LRRK2. ( C ) Western blot analysis of Rab8a and phosphorylated Rab8a (pT72-Rab8a) in KD and OE compared with WT cells. Top, representative blot; bottom, densitometric quantification normalized to actin. ( D ) Representative super-resolution immunofluorescence images of LAMP1 (magenta) and LRRK2 (green) in WT ( D1 ), NAPE-PLD KD ( D2 ), and PLA2G4E OE ( D3 ) cells. Nuclei were stained with DAPI (gray). Scale bar, 5 μm. ( E ) Fluorescence intensity quantification and representative images of the Magic Red–cathepsin B assay, indicating lysosomal degradative capacity per cell, in WT ( E1 ), NAPE-PLD KD ( E2 ), and PLA2G4E OE ( E3 ) cells. Representative images are displayed using the FIRE lookup table (LUT) to indicate fluorescence intensity. Scale bars, 10 μm. Mean values are indicated in the graphs; error bars represent SEM. *P < 0.05, ****P < 0.0001; one-way ANOVA with Tukey’s multiple-comparison test.

    Article Snippet: The NAPE-PLD Human Gene Knockout Kit (CRISPR, Origene, KN409877) was used following manufacturer’s instructions.

    Techniques: Immunofluorescence, Knockdown, Western Blot, Staining, Fluorescence, Comparison

    ( A ) Immunofluorescence analysis of LRRK2 (gray) with representative images ( A1-A4 ) and quantification in WT ( A1, A2 ), NAPE-PLD-knockdown (KD; A3 ), and PLA2G4E-overexpressing (OE; A4 ) cells treated or not with α-syn ( A2-A4 ). ( B ) Quantification of the colocalization between LAMP1-positive structures and LRRK2, expressed as Mander’s overlap coefficient for the fraction of LAMP1 signal overlapping with LRRK2. ( C ) Quantification and representative images of α-syn aggregate number per cell in WT ( C1 ), NAPE-PLD KD ( C2 ) and PLA2G4E OE ( C3 ) cells. LAMP1 is shown in yellow, α-syn in pink, and nuclei were counterstained with DAPI (blue). Scale bar, 10 μm. Mean values are indicated in the graphs; error bars represent SEM. ***P < 0.001, ****P < 0.0001; one-way ANOVA with Tuckey’s multiple-comparison test.

    Journal: bioRxiv

    Article Title: A NAPE–LRRK2 metabolic axis controls lysosomal homeostasis in Parkinson’s disease

    doi: 10.64898/2026.03.19.712902

    Figure Lengend Snippet: ( A ) Immunofluorescence analysis of LRRK2 (gray) with representative images ( A1-A4 ) and quantification in WT ( A1, A2 ), NAPE-PLD-knockdown (KD; A3 ), and PLA2G4E-overexpressing (OE; A4 ) cells treated or not with α-syn ( A2-A4 ). ( B ) Quantification of the colocalization between LAMP1-positive structures and LRRK2, expressed as Mander’s overlap coefficient for the fraction of LAMP1 signal overlapping with LRRK2. ( C ) Quantification and representative images of α-syn aggregate number per cell in WT ( C1 ), NAPE-PLD KD ( C2 ) and PLA2G4E OE ( C3 ) cells. LAMP1 is shown in yellow, α-syn in pink, and nuclei were counterstained with DAPI (blue). Scale bar, 10 μm. Mean values are indicated in the graphs; error bars represent SEM. ***P < 0.001, ****P < 0.0001; one-way ANOVA with Tuckey’s multiple-comparison test.

    Article Snippet: The NAPE-PLD Human Gene Knockout Kit (CRISPR, Origene, KN409877) was used following manufacturer’s instructions.

    Techniques: Immunofluorescence, Knockdown, Comparison

    ( A ) Schematic representation of the generation of stable SH-SY5Y cell lines overexpressing NAPE-PLD, leading to reduced NAPE levels at cellular membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-overexpressing (OE; green) and wild-type (WT; blue) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. ( E ) Immunofluorescence analysis of LRRK2 (gray) with representative images ( E1, E2 ) and quantification in WT ( E1 ) and NAPE-PLD OE ( E2 ) cells. Nuclei were counterstained with DAPI (blue). ( F ) Western blot analysis of Rab8a and phosphorylated Rab8a (pT72-Rab8a) in NAPE-PLD OE and WT cells. Top: representative blot; bottom, densitometric quantification normalized to actin. ( G ) Schematic representation of the experimental strategy. Ionomycin treatment induces intracellular Ca 2+ release, promoting NAPE production and degradation in NAPE-PLD OE cells. NAPE-PLD activity was inhibited by overnight treatment with the NAPE-PLD inhibitor LEI-401. Ionomycin was administered together with Magic Red reagent for 30 min before image acquisition. ( H ) Fluorescence intensity quantification and representative images of the Magic Red – cathepsin B assay, indicating lysosomal degradative capacity per cell, in untreated WT ( H1, white bar and blue dots) and NAPE-PLD OE cells ( H4, white bar and green dots), ionomycin-treated WT ( H2, blue bar and blue dots) and NAPE-PLD OE cells ( H5, green bar and green dots) or WT ( H3, blue bar white dots) and NAPE-PLD OE cells ( H6 , green bar and white dots) treated with ionomycin and LEI-401. Representative images are displayed using the FIRE lookup table (LUT) to indicate fluorescence intensity. Scale bars, 10 μm. Mean values are indicated in the graphs; error bars represent SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; Student’s t -test with Welch’s correction ( B-F ) and two-way ANOVA with Tuckey’s multiple-comparisons test ( H ).

    Journal: bioRxiv

    Article Title: A NAPE–LRRK2 metabolic axis controls lysosomal homeostasis in Parkinson’s disease

    doi: 10.64898/2026.03.19.712902

    Figure Lengend Snippet: ( A ) Schematic representation of the generation of stable SH-SY5Y cell lines overexpressing NAPE-PLD, leading to reduced NAPE levels at cellular membranes. ( B ) NAPE-PLD mRNA levels in NAPE-PLD-overexpressing (OE; green) and wild-type (WT; blue) cells. ( C ) Western blot analysis of NAPE-PLD protein levels in OE and WT cells. Top, representative blot; bottom, densitometric quantification normalized to GAPDH. ( D ) Immunofluorescence analysis of NAPE-PLD (gray) with representative images ( D1, D2 ) and quantification. ( E ) Immunofluorescence analysis of LRRK2 (gray) with representative images ( E1, E2 ) and quantification in WT ( E1 ) and NAPE-PLD OE ( E2 ) cells. Nuclei were counterstained with DAPI (blue). ( F ) Western blot analysis of Rab8a and phosphorylated Rab8a (pT72-Rab8a) in NAPE-PLD OE and WT cells. Top: representative blot; bottom, densitometric quantification normalized to actin. ( G ) Schematic representation of the experimental strategy. Ionomycin treatment induces intracellular Ca 2+ release, promoting NAPE production and degradation in NAPE-PLD OE cells. NAPE-PLD activity was inhibited by overnight treatment with the NAPE-PLD inhibitor LEI-401. Ionomycin was administered together with Magic Red reagent for 30 min before image acquisition. ( H ) Fluorescence intensity quantification and representative images of the Magic Red – cathepsin B assay, indicating lysosomal degradative capacity per cell, in untreated WT ( H1, white bar and blue dots) and NAPE-PLD OE cells ( H4, white bar and green dots), ionomycin-treated WT ( H2, blue bar and blue dots) and NAPE-PLD OE cells ( H5, green bar and green dots) or WT ( H3, blue bar white dots) and NAPE-PLD OE cells ( H6 , green bar and white dots) treated with ionomycin and LEI-401. Representative images are displayed using the FIRE lookup table (LUT) to indicate fluorescence intensity. Scale bars, 10 μm. Mean values are indicated in the graphs; error bars represent SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; Student’s t -test with Welch’s correction ( B-F ) and two-way ANOVA with Tuckey’s multiple-comparisons test ( H ).

    Article Snippet: The NAPE-PLD Human Gene Knockout Kit (CRISPR, Origene, KN409877) was used following manufacturer’s instructions.

    Techniques: Western Blot, Immunofluorescence, Activity Assay, Fluorescence

    ( A ) Immunofluorescence analysis of LRRK2 (gray) with representative images ( A1-A3 ) and quantification in WT ( A1, A2 ), and NAPE-PLD OE ( A3 ) SH-SY5Y cells treated or not with α-synuclein (α-syn; A2, A3 ). ( B ) Quantification and representative images of α-syn aggregate number per cell in WT ( B1 ) and NAPE-PLD OE ( B2 ) cells. α-Syn is shown in pink, and nuclei were counterstained with DAPI. Scale bar, 10 μm. Mean values are indicated in the graphs; error bars represent SEM. *P<0.05, ***P < 0.001, ****P < 0.0001; one-way ANOVA with Tuckey’s multiple-comparison test ( A ) and Student’s t -test with Welch’s correction ( B ).

    Journal: bioRxiv

    Article Title: A NAPE–LRRK2 metabolic axis controls lysosomal homeostasis in Parkinson’s disease

    doi: 10.64898/2026.03.19.712902

    Figure Lengend Snippet: ( A ) Immunofluorescence analysis of LRRK2 (gray) with representative images ( A1-A3 ) and quantification in WT ( A1, A2 ), and NAPE-PLD OE ( A3 ) SH-SY5Y cells treated or not with α-synuclein (α-syn; A2, A3 ). ( B ) Quantification and representative images of α-syn aggregate number per cell in WT ( B1 ) and NAPE-PLD OE ( B2 ) cells. α-Syn is shown in pink, and nuclei were counterstained with DAPI. Scale bar, 10 μm. Mean values are indicated in the graphs; error bars represent SEM. *P<0.05, ***P < 0.001, ****P < 0.0001; one-way ANOVA with Tuckey’s multiple-comparison test ( A ) and Student’s t -test with Welch’s correction ( B ).

    Article Snippet: The NAPE-PLD Human Gene Knockout Kit (CRISPR, Origene, KN409877) was used following manufacturer’s instructions.

    Techniques: Immunofluorescence, Comparison

    ( A ) Representative images of the Magic Red – cathepsin B assay, indicating lysosomal degradative capacity per cell, in healthy dopaminergic neurons (DANs; A1, A2 ) and G2019S-LRRK2 mutant DANs ( A3, A4 ) treated ( A2, A4 ) or not ( A1, A3 ) with the NAPE-PLD inhibitor LEI-401. ( B ) Fluorescence intensity quantification of the Magic Red assay in healthy DANs (green) and G2019S-LRRK2 mutant DANs (gray). ( C, D ) Quantification and representative images of α-synuclein (α-syn) aggregate number per cell in healthy DANs ( D1, D2 ) and G2019-LRRK2 mutant DANs ( D3, D4 ) treated ( D2, D4 ) or not ( D1, D3 ) with LEI-401. α-Syn is shown in pink, LAMP1 yellow, and nuclei were counterstained with DAPI. Scale bars, 10 μm. *P < 0.05, ****P < 0.0001; two-way ANOVA with Tuckey’s multiple-comparison test.

    Journal: bioRxiv

    Article Title: A NAPE–LRRK2 metabolic axis controls lysosomal homeostasis in Parkinson’s disease

    doi: 10.64898/2026.03.19.712902

    Figure Lengend Snippet: ( A ) Representative images of the Magic Red – cathepsin B assay, indicating lysosomal degradative capacity per cell, in healthy dopaminergic neurons (DANs; A1, A2 ) and G2019S-LRRK2 mutant DANs ( A3, A4 ) treated ( A2, A4 ) or not ( A1, A3 ) with the NAPE-PLD inhibitor LEI-401. ( B ) Fluorescence intensity quantification of the Magic Red assay in healthy DANs (green) and G2019S-LRRK2 mutant DANs (gray). ( C, D ) Quantification and representative images of α-synuclein (α-syn) aggregate number per cell in healthy DANs ( D1, D2 ) and G2019-LRRK2 mutant DANs ( D3, D4 ) treated ( D2, D4 ) or not ( D1, D3 ) with LEI-401. α-Syn is shown in pink, LAMP1 yellow, and nuclei were counterstained with DAPI. Scale bars, 10 μm. *P < 0.05, ****P < 0.0001; two-way ANOVA with Tuckey’s multiple-comparison test.

    Article Snippet: The NAPE-PLD Human Gene Knockout Kit (CRISPR, Origene, KN409877) was used following manufacturer’s instructions.

    Techniques: Mutagenesis, Fluorescence, Comparison