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anti pea15  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti pea15
    Anti Pea15, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+pea15/pmc13058230-102-7-10
    Average 86 stars, based on 1 article reviews
    anti pea15 - by Bioz Stars, 2026-10
    86/100 stars

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    Related Articles

    Purification:

    Article Title: FADD adaptor and PEA-15/ERK1/2 partners in major depression and schizophrenia postmortem brains: basal contents and effects of psychotropic treatments.
    Article Snippet: Enhanced brain apoptosis (neurons and glia) may be involved in major depression (MD) and schizophrenia (SZ), mainly through the activation of the intrinsic (mitochondrial) apoptotic pathway.. In the extrinsic death pathway, pro-apoptotic Fas-associated death domain (FADD) adaptor and its non-apoptotic p-Ser194 FADD form have critical roles interacting with other death regulators such as phosphoprotein enriched in astrocytes of 15 kDa (PEA-15) and extracellular signal-regulated kinase (ERK).. The basal status of FADD (protein and messenger RNA (mRNA)) and the effects of psychotropic drugs (detected in blood/urine samples) were first assessed in postmortem prefrontal cortex of MD and SZ subjects (including a non-MD/SZ suicide group).

    Bioprocessing:

    Article Title: FADD adaptor and PEA-15/ERK1/2 partners in major depression and schizophrenia postmortem brains: basal contents and effects of psychotropic treatments.
    Article Snippet: Enhanced brain apoptosis (neurons and glia) may be involved in major depression (MD) and schizophrenia (SZ), mainly through the activation of the intrinsic (mitochondrial) apoptotic pathway.. In the extrinsic death pathway, pro-apoptotic Fas-associated death domain (FADD) adaptor and its non-apoptotic p-Ser194 FADD form have critical roles interacting with other death regulators such as phosphoprotein enriched in astrocytes of 15 kDa (PEA-15) and extracellular signal-regulated kinase (ERK).. The basal status of FADD (protein and messenger RNA (mRNA)) and the effects of psychotropic drugs (detected in blood/urine samples) were first assessed in postmortem prefrontal cortex of MD and SZ subjects (including a non-MD/SZ suicide group).

    Membrane:

    Article Title: Overexpression of phosphoprotein enriched in astrocytes 15 reverses the damage induced by propofol in hippocampal neurons
    Article Snippet: .. The PVDF membrane was sealed and blocked using 5% non-fat milk at 37°C for 1.5 h. Subsequently, the PVDF membrane was incubated with primary antibodies at 4°C for 24 h. The following antibodies were used in the present study: Anti-PEA15 [1:800; Cell Signaling Technology (CST), Inc., Danvers, MA, USA; cat. no. 2780], anti-pro-caspase-3 (1:1,000; Abcam, Cambridge, UK; cat. no. ab32150), anti-active caspase-3 (1:600; Abcam; cat. no. ab13847), anti-B-cell lymphoma-2 (Bcl-2; 1:700; Abcam; cat. no. ab59348), anti-apoptosis regulator BAX (Bax; 1:1,000; Abcam; cat. no. ab32503), anti-phosphorylated (p)-extracellular signal-regulated kinases 1/2 (ERK1/2; 1:1,000; CST, Inc.; cat. no. 4370), anti-ERK1/2 (1:600; CST, Inc.; cat. no. 9102), anti-ribosomal S6 kinase 2 (RSK2; 1:500; Abcam; cat. no. ab32133), anti-p-cAMP-response element binding protein 1 (CREB1; 1:800; Abcam; cat. no. ab32096), anti-CREB1 (1:700; Abcam; cat. no. ab32515) and anti-β-actin (1:5,000; Abcam; cat. no. ab179467). .. The PVDF membrane was subsequently incubated with Horseradish peroxidase-conjugated secondary antibodies (goat anti-rabbit; cat. no. ab205718; 1:2,000) at room temperature for 1 h. The proteins were detected using enhanced chemiluminescent reagents (Thermo Fisher Scientific, Inc.).

    Article Title: Overexpression of phosphoprotein enriched in astrocytes 15 reverses the damage induced by propofol in hippocampal neurons.
    Article Snippet: .. The PVDF membrane was sealed and blocked using 5% non‐fat milk at 37 ̊C for 1.5 h. Subsequently, the PVDF membrane was incubated with primary antibodies at 4 ̊C for 24 h. The following antibodies were used in the present study: Anti‐PEA15 [1:800; Cell Signaling Technology (CST), Inc., Danvers, MA, USA; cat. no. 2780], anti‐pro‐caspase‐3 (1:1,000; Abcam, Cambridge, UK; cat. no. ab32150), anti‐active caspase‐3 (1:600; Abcam; cat. no. ab13847), anti‐B‐cell lymphoma‐2 (Bcl‐2; 1:700; Abcam; cat. no. ab59348), anti‐apoptosis regulator BAX (Bax; 1:1,000; Abcam; cat. no. ab32503), anti‐phosphorylated (p)-extracellular signal‐regulated kinases 1/2 (erK1/2; 1:1,000; CST, Inc.; cat. no. 4370), anti‐ERK1/2 (1:600; CST, Inc.; cat. no. 9102), anti‐ribosomal S6 kinase 2 (RSK2; 1:500; Abcam; cat. no. ab32133), anti‐p‐cAMP‐response element binding protein 1 (CREB1; 1:800; Abcam; cat. no. ab32096), anti‐CREB1 (1:700; Abcam; cat. no. ab32515) and anti-β‐actin (1:5,000; Abcam; cat. no. ab179467). .. The PVDF membrane was subsequently incubated with Horseradish peroxidase‐conjugated secondary antibodies (goat anti‐rabbit; cat. no. ab205718; 1:2,000) at room temperature for 1 h. The proteins were detected using enhanced chemiluminescent reagents (Thermo Fisher Scientific, Inc.).

    Incubation:

    Article Title: Overexpression of phosphoprotein enriched in astrocytes 15 reverses the damage induced by propofol in hippocampal neurons
    Article Snippet: .. The PVDF membrane was sealed and blocked using 5% non-fat milk at 37°C for 1.5 h. Subsequently, the PVDF membrane was incubated with primary antibodies at 4°C for 24 h. The following antibodies were used in the present study: Anti-PEA15 [1:800; Cell Signaling Technology (CST), Inc., Danvers, MA, USA; cat. no. 2780], anti-pro-caspase-3 (1:1,000; Abcam, Cambridge, UK; cat. no. ab32150), anti-active caspase-3 (1:600; Abcam; cat. no. ab13847), anti-B-cell lymphoma-2 (Bcl-2; 1:700; Abcam; cat. no. ab59348), anti-apoptosis regulator BAX (Bax; 1:1,000; Abcam; cat. no. ab32503), anti-phosphorylated (p)-extracellular signal-regulated kinases 1/2 (ERK1/2; 1:1,000; CST, Inc.; cat. no. 4370), anti-ERK1/2 (1:600; CST, Inc.; cat. no. 9102), anti-ribosomal S6 kinase 2 (RSK2; 1:500; Abcam; cat. no. ab32133), anti-p-cAMP-response element binding protein 1 (CREB1; 1:800; Abcam; cat. no. ab32096), anti-CREB1 (1:700; Abcam; cat. no. ab32515) and anti-β-actin (1:5,000; Abcam; cat. no. ab179467). .. The PVDF membrane was subsequently incubated with Horseradish peroxidase-conjugated secondary antibodies (goat anti-rabbit; cat. no. ab205718; 1:2,000) at room temperature for 1 h. The proteins were detected using enhanced chemiluminescent reagents (Thermo Fisher Scientific, Inc.).

    Article Title: Secretome protein signature of human gastrointestinal stromal tumor cells.
    Article Snippet: Strategies for correct diagnosis, treatment evaluation and recurrence prediction are important for the prognosis and mortality rates among cancer patients.. In spite of major improvements in clinical management, gastrointestinal stromal tumors (GISTs) can still be deadly due to metastasis and recurrences, which confirms the unmet need of reliable follow-up modalities.. Tumor-specific secreted, shed or leaked proteins (collectively known as secretome) are considered promising sources for biomarkers, and suitable for detection in biofluids.

    Article Title: Overexpression of phosphoprotein enriched in astrocytes 15 reverses the damage induced by propofol in hippocampal neurons.
    Article Snippet: .. The PVDF membrane was sealed and blocked using 5% non‐fat milk at 37 ̊C for 1.5 h. Subsequently, the PVDF membrane was incubated with primary antibodies at 4 ̊C for 24 h. The following antibodies were used in the present study: Anti‐PEA15 [1:800; Cell Signaling Technology (CST), Inc., Danvers, MA, USA; cat. no. 2780], anti‐pro‐caspase‐3 (1:1,000; Abcam, Cambridge, UK; cat. no. ab32150), anti‐active caspase‐3 (1:600; Abcam; cat. no. ab13847), anti‐B‐cell lymphoma‐2 (Bcl‐2; 1:700; Abcam; cat. no. ab59348), anti‐apoptosis regulator BAX (Bax; 1:1,000; Abcam; cat. no. ab32503), anti‐phosphorylated (p)-extracellular signal‐regulated kinases 1/2 (erK1/2; 1:1,000; CST, Inc.; cat. no. 4370), anti‐ERK1/2 (1:600; CST, Inc.; cat. no. 9102), anti‐ribosomal S6 kinase 2 (RSK2; 1:500; Abcam; cat. no. ab32133), anti‐p‐cAMP‐response element binding protein 1 (CREB1; 1:800; Abcam; cat. no. ab32096), anti‐CREB1 (1:700; Abcam; cat. no. ab32515) and anti-β‐actin (1:5,000; Abcam; cat. no. ab179467). .. The PVDF membrane was subsequently incubated with Horseradish peroxidase‐conjugated secondary antibodies (goat anti‐rabbit; cat. no. ab205718; 1:2,000) at room temperature for 1 h. The proteins were detected using enhanced chemiluminescent reagents (Thermo Fisher Scientific, Inc.).

    Binding Assay:

    Article Title: Overexpression of phosphoprotein enriched in astrocytes 15 reverses the damage induced by propofol in hippocampal neurons
    Article Snippet: .. The PVDF membrane was sealed and blocked using 5% non-fat milk at 37°C for 1.5 h. Subsequently, the PVDF membrane was incubated with primary antibodies at 4°C for 24 h. The following antibodies were used in the present study: Anti-PEA15 [1:800; Cell Signaling Technology (CST), Inc., Danvers, MA, USA; cat. no. 2780], anti-pro-caspase-3 (1:1,000; Abcam, Cambridge, UK; cat. no. ab32150), anti-active caspase-3 (1:600; Abcam; cat. no. ab13847), anti-B-cell lymphoma-2 (Bcl-2; 1:700; Abcam; cat. no. ab59348), anti-apoptosis regulator BAX (Bax; 1:1,000; Abcam; cat. no. ab32503), anti-phosphorylated (p)-extracellular signal-regulated kinases 1/2 (ERK1/2; 1:1,000; CST, Inc.; cat. no. 4370), anti-ERK1/2 (1:600; CST, Inc.; cat. no. 9102), anti-ribosomal S6 kinase 2 (RSK2; 1:500; Abcam; cat. no. ab32133), anti-p-cAMP-response element binding protein 1 (CREB1; 1:800; Abcam; cat. no. ab32096), anti-CREB1 (1:700; Abcam; cat. no. ab32515) and anti-β-actin (1:5,000; Abcam; cat. no. ab179467). .. The PVDF membrane was subsequently incubated with Horseradish peroxidase-conjugated secondary antibodies (goat anti-rabbit; cat. no. ab205718; 1:2,000) at room temperature for 1 h. The proteins were detected using enhanced chemiluminescent reagents (Thermo Fisher Scientific, Inc.).

    Article Title: Overexpression of phosphoprotein enriched in astrocytes 15 reverses the damage induced by propofol in hippocampal neurons.
    Article Snippet: .. The PVDF membrane was sealed and blocked using 5% non‐fat milk at 37 ̊C for 1.5 h. Subsequently, the PVDF membrane was incubated with primary antibodies at 4 ̊C for 24 h. The following antibodies were used in the present study: Anti‐PEA15 [1:800; Cell Signaling Technology (CST), Inc., Danvers, MA, USA; cat. no. 2780], anti‐pro‐caspase‐3 (1:1,000; Abcam, Cambridge, UK; cat. no. ab32150), anti‐active caspase‐3 (1:600; Abcam; cat. no. ab13847), anti‐B‐cell lymphoma‐2 (Bcl‐2; 1:700; Abcam; cat. no. ab59348), anti‐apoptosis regulator BAX (Bax; 1:1,000; Abcam; cat. no. ab32503), anti‐phosphorylated (p)-extracellular signal‐regulated kinases 1/2 (erK1/2; 1:1,000; CST, Inc.; cat. no. 4370), anti‐ERK1/2 (1:600; CST, Inc.; cat. no. 9102), anti‐ribosomal S6 kinase 2 (RSK2; 1:500; Abcam; cat. no. ab32133), anti‐p‐cAMP‐response element binding protein 1 (CREB1; 1:800; Abcam; cat. no. ab32096), anti‐CREB1 (1:700; Abcam; cat. no. ab32515) and anti-β‐actin (1:5,000; Abcam; cat. no. ab179467). .. The PVDF membrane was subsequently incubated with Horseradish peroxidase‐conjugated secondary antibodies (goat anti‐rabbit; cat. no. ab205718; 1:2,000) at room temperature for 1 h. The proteins were detected using enhanced chemiluminescent reagents (Thermo Fisher Scientific, Inc.).

    Blocking Assay:

    Article Title: Secretome protein signature of human gastrointestinal stromal tumor cells.
    Article Snippet: Strategies for correct diagnosis, treatment evaluation and recurrence prediction are important for the prognosis and mortality rates among cancer patients.. In spite of major improvements in clinical management, gastrointestinal stromal tumors (GISTs) can still be deadly due to metastasis and recurrences, which confirms the unmet need of reliable follow-up modalities.. Tumor-specific secreted, shed or leaked proteins (collectively known as secretome) are considered promising sources for biomarkers, and suitable for detection in biofluids.

    Transduction:

    Article Title: PLD1 participates in BDNF-induced signalling in cortical neurons
    Article Snippet: .. Antibodies anti-HA (Babco), anti-RSK2, anti-APPL1, anti-Rab7, anti-TrkB (Santa Cruz Bio-technology), anti-PLD1, anti-ERK, anti-phospho-ERK, (New England BioLabs), anti-β-tubulin, anti-CREB (Millipore), anti-GAPDH, anti-phospho-CREB (Ser-133), anti-mTOR, anti-phospho-mTOR (Ser-2481), anti-phospho-S6K (Thr-389), anti-phospho-S6K (Thr-421/Ser-424), anti-PEA15 (Cell Signalling), anti-Rab5 (Transduction Laboratories) were used. ..

    Western Blot:

    Article Title: Proliferation and Apoptosis Adaptor Protein 15 (PEA15), a Potential Oncogenic Regulator of VHL and HIF1A Identified through Proteomic Analysis in Hepatocellular Carcinoma
    Article Snippet: .. For WB, the following antibodies were used: anti-PEA15 (1:500, #13091, Cell Signaling Technology), anti-PEA15 (1:500, sc-166678, Santa Cruz Biotechnology), anti-phospho-PEA15 (Ser 104 , 1:1,000, #2776, Cell Signaling Technology), anti-phospho-PEA15 (Ser 116 , 1:1,000, #44-836G, Invitrogen), anti-VHL (1: 5,000, sc-135657, Santa Cruz Biotechnology), anti-β-actin (1:5,000, sc-4778, Santa Cruz Biotechnology), anti-α-tubulin (1:5,000, sc-32293, Santa Cruz Biotechnology), anti-HIF1A (1:500, #610958, BD Biosciences), anti-prolyl hydroxylase domain containing protein 2 (PHD2) (1:1,000, sc-271835, Santa Cruz Biotechnology), anti-factor inhibiting HIF-1 (FIH1) (1:1,000, sc-365128, Santa Cruz Biotechnology), anti-glutathione S -transferase (GST) (1:5,000, sc-138, Santa Cruz Biotechnology), anti-hemagglutinin tag (HA) (1:5,000, #901513, BioLegend), anti-histidine (His) probe (1:1,000, sc-8036, Santa Cruz Biotechnology), anti-elongin B (ELOB) (1:1,000, #629302, BioLegend), anti-elongin C (ELOC) (1:1,000, #613101, BioLegend), anti-cullin 2 (1:1,000, #51-1800, Invitrogen), anti-ring box 1 (RBX1) (1:1,000, 11922, Cell Signaling Technology), anti-Snail family transcriptional repressor 1 (SNAI1) (1:1,000, #3879, Cell Signaling Technology), anti-signal transducer and activator of transcription 3 (STAT3) (1:1,000, #9139, Cell Signaling Technology), anti-SMAD family member 3 (SMAD3) (1:1,000, #9523, Cell Signaling Technology), and anti-vascular endothelial growth factor A (VEGFA) (1:1,000, #50661, Cell Signaling Technology). .. Horseradish peroxidase-linked anti-rabbit IgG (7074S, Cell Signaling Technology), horseradish peroxidase-linked anti-mouse IgG (7076S, Cell Signaling Technology), Mouse TrueBlot ULTRA (18-8817-33, Rockland Immunochemicals), and Rabbit TrueBlot (18-8816-33, Rockland Immunochemicals) were used as secondary antibodies for WB.



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    Clinical characteristics and prognostic factor <t>PEA15</t> in the OS autophagy-related model (A) Kaplan-Meier survival analysis of the OS prognostic factor PEA15. (B) Risk survival status plot. (C and D) Univariate and multivariate COX regression analyses of the OS autophagy-related model and various clinical features. (E) mRNA expression levels of PEA15 in the normal osteoblast cell line hFOB1.19 and four osteosarcoma cell lines. (F and G) Validation of knockdown efficiency in U2OS cells and overexpression efficiency in MG63 cells ( n = 3). (H and I) CCK-8 assays of cell proliferation following PEA15 knockdown and overexpression ( n = 3). (J–M) Validation of autophagy activity in human tumor tissues and corresponding statistical analysis ( n = 3). (N and O) Protein expression levels of PEA15 are validated by knockdown and overexpression experiments. The data are presented as mean ± SD. ∗∗ p < 0.01, ∗∗∗ p < 0.001. n.s.: not significant.
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    Image Search Results


    Clinical characteristics and prognostic factor PEA15 in the OS autophagy-related model (A) Kaplan-Meier survival analysis of the OS prognostic factor PEA15. (B) Risk survival status plot. (C and D) Univariate and multivariate COX regression analyses of the OS autophagy-related model and various clinical features. (E) mRNA expression levels of PEA15 in the normal osteoblast cell line hFOB1.19 and four osteosarcoma cell lines. (F and G) Validation of knockdown efficiency in U2OS cells and overexpression efficiency in MG63 cells ( n = 3). (H and I) CCK-8 assays of cell proliferation following PEA15 knockdown and overexpression ( n = 3). (J–M) Validation of autophagy activity in human tumor tissues and corresponding statistical analysis ( n = 3). (N and O) Protein expression levels of PEA15 are validated by knockdown and overexpression experiments. The data are presented as mean ± SD. ∗∗ p < 0.01, ∗∗∗ p < 0.001. n.s.: not significant.

    Journal: iScience

    Article Title: PEA15 promotes osteosarcoma progression and cisplatin resistance by modulating autophagy through the FABP3-TNF signaling axis

    doi: 10.1016/j.isci.2025.113695

    Figure Lengend Snippet: Clinical characteristics and prognostic factor PEA15 in the OS autophagy-related model (A) Kaplan-Meier survival analysis of the OS prognostic factor PEA15. (B) Risk survival status plot. (C and D) Univariate and multivariate COX regression analyses of the OS autophagy-related model and various clinical features. (E) mRNA expression levels of PEA15 in the normal osteoblast cell line hFOB1.19 and four osteosarcoma cell lines. (F and G) Validation of knockdown efficiency in U2OS cells and overexpression efficiency in MG63 cells ( n = 3). (H and I) CCK-8 assays of cell proliferation following PEA15 knockdown and overexpression ( n = 3). (J–M) Validation of autophagy activity in human tumor tissues and corresponding statistical analysis ( n = 3). (N and O) Protein expression levels of PEA15 are validated by knockdown and overexpression experiments. The data are presented as mean ± SD. ∗∗ p < 0.01, ∗∗∗ p < 0.001. n.s.: not significant.

    Article Snippet: The membrane was blocked with 5% non-fat milk in TBST for 2 h at room temperature, followed by incubation overnight at 4°C with primary antibodies targeting PEA15 (CST, #13091,1:1000), LC3B (CST, #3868,1:1000), ATG5 (CST,#9980,1:1000),Cleaved-Caspase3 (CST,#9664,1:1000),Cleaved-PARP (CST, #9541,1:1000), FABP3 (CST,#14780,1:1000), TNF-α (CST,#3707,1:1000), and GAPDH (CST, #5174,1:1000).

    Techniques: Expressing, Biomarker Discovery, Knockdown, Over Expression, CCK-8 Assay, Activity Assay

    PEA15 influences tumor cell growth (A and B) Wound healing assays to evaluate the effects of PEA15 knockdown and overexpression. (C and D) Colony formation assays to assess the impact of PEA15 knockdown and overexpression. (E–G) Migration and invasion assays in U2OS cells with PEA15 knockdown, including statistical analysis and expression levels of migration- and invasion-related proteins ( n = 3). (H–J) Migration and invasion assays in MG63 cells with PEA15 overexpression, along with the corresponding protein expression profiles ( n = 3). The data are presented as mean ± SD. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

    Journal: iScience

    Article Title: PEA15 promotes osteosarcoma progression and cisplatin resistance by modulating autophagy through the FABP3-TNF signaling axis

    doi: 10.1016/j.isci.2025.113695

    Figure Lengend Snippet: PEA15 influences tumor cell growth (A and B) Wound healing assays to evaluate the effects of PEA15 knockdown and overexpression. (C and D) Colony formation assays to assess the impact of PEA15 knockdown and overexpression. (E–G) Migration and invasion assays in U2OS cells with PEA15 knockdown, including statistical analysis and expression levels of migration- and invasion-related proteins ( n = 3). (H–J) Migration and invasion assays in MG63 cells with PEA15 overexpression, along with the corresponding protein expression profiles ( n = 3). The data are presented as mean ± SD. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

    Article Snippet: The membrane was blocked with 5% non-fat milk in TBST for 2 h at room temperature, followed by incubation overnight at 4°C with primary antibodies targeting PEA15 (CST, #13091,1:1000), LC3B (CST, #3868,1:1000), ATG5 (CST,#9980,1:1000),Cleaved-Caspase3 (CST,#9664,1:1000),Cleaved-PARP (CST, #9541,1:1000), FABP3 (CST,#14780,1:1000), TNF-α (CST,#3707,1:1000), and GAPDH (CST, #5174,1:1000).

    Techniques: Knockdown, Over Expression, Migration, Expressing

    PEA15 modulates autophagy and apoptosis (A and B) Assessment of autophagic activity (autophagosome formation) following PEA15 knockdown and overexpression ( n = 3). (C and D) Apoptosis assays were carried out to evaluate the effects of PEA15 knockdown and overexpression ( n = 3). (E and F) Impact of PEA15 knockdown and overexpression on the expression of autophagy-related proteins. (G and H) Changes in apoptosis-related protein levels after PEA15 knockdown and overexpression. The data are presented as mean ± SD. ∗∗ p < 0.01.

    Journal: iScience

    Article Title: PEA15 promotes osteosarcoma progression and cisplatin resistance by modulating autophagy through the FABP3-TNF signaling axis

    doi: 10.1016/j.isci.2025.113695

    Figure Lengend Snippet: PEA15 modulates autophagy and apoptosis (A and B) Assessment of autophagic activity (autophagosome formation) following PEA15 knockdown and overexpression ( n = 3). (C and D) Apoptosis assays were carried out to evaluate the effects of PEA15 knockdown and overexpression ( n = 3). (E and F) Impact of PEA15 knockdown and overexpression on the expression of autophagy-related proteins. (G and H) Changes in apoptosis-related protein levels after PEA15 knockdown and overexpression. The data are presented as mean ± SD. ∗∗ p < 0.01.

    Article Snippet: The membrane was blocked with 5% non-fat milk in TBST for 2 h at room temperature, followed by incubation overnight at 4°C with primary antibodies targeting PEA15 (CST, #13091,1:1000), LC3B (CST, #3868,1:1000), ATG5 (CST,#9980,1:1000),Cleaved-Caspase3 (CST,#9664,1:1000),Cleaved-PARP (CST, #9541,1:1000), FABP3 (CST,#14780,1:1000), TNF-α (CST,#3707,1:1000), and GAPDH (CST, #5174,1:1000).

    Techniques: Activity Assay, Knockdown, Over Expression, Expressing

    Mechanistic insights into PEA15 downstream signaling (A) Heatmap of transcriptomic analysis in U2OS cells following PEA15 knockdown. (B) CCK-8 assay was adopted to assess cell proliferation after FABP3 knockdown in U2OS cells ( n = 3). (C) Colony formation assay in U2OS cells with FABP3 knockdown ( n = 3). (D and E) Migration and invasion assays in U2OS cells with FABP3 knockdown, along with the expression levels of related proteins ( n = 3). (F and G) Fluorescence imaging of autophagic activity and expression of autophagy-related proteins following FABP3 knockdown. (K–L) Flow cytometry analysis of apoptosis and expression of apoptosis-related markers after FABP3 knockdown ( n = 3). (M) KEGG pathway analysis of transcriptomic data from U2OS cells with PEA15 knockdown. (N) In U2OS cells with PEA15 knockdown, FABP3 was overexpressed, followed by the detection of ATG5 and LC3B protein expression levels.

    Journal: iScience

    Article Title: PEA15 promotes osteosarcoma progression and cisplatin resistance by modulating autophagy through the FABP3-TNF signaling axis

    doi: 10.1016/j.isci.2025.113695

    Figure Lengend Snippet: Mechanistic insights into PEA15 downstream signaling (A) Heatmap of transcriptomic analysis in U2OS cells following PEA15 knockdown. (B) CCK-8 assay was adopted to assess cell proliferation after FABP3 knockdown in U2OS cells ( n = 3). (C) Colony formation assay in U2OS cells with FABP3 knockdown ( n = 3). (D and E) Migration and invasion assays in U2OS cells with FABP3 knockdown, along with the expression levels of related proteins ( n = 3). (F and G) Fluorescence imaging of autophagic activity and expression of autophagy-related proteins following FABP3 knockdown. (K–L) Flow cytometry analysis of apoptosis and expression of apoptosis-related markers after FABP3 knockdown ( n = 3). (M) KEGG pathway analysis of transcriptomic data from U2OS cells with PEA15 knockdown. (N) In U2OS cells with PEA15 knockdown, FABP3 was overexpressed, followed by the detection of ATG5 and LC3B protein expression levels.

    Article Snippet: The membrane was blocked with 5% non-fat milk in TBST for 2 h at room temperature, followed by incubation overnight at 4°C with primary antibodies targeting PEA15 (CST, #13091,1:1000), LC3B (CST, #3868,1:1000), ATG5 (CST,#9980,1:1000),Cleaved-Caspase3 (CST,#9664,1:1000),Cleaved-PARP (CST, #9541,1:1000), FABP3 (CST,#14780,1:1000), TNF-α (CST,#3707,1:1000), and GAPDH (CST, #5174,1:1000).

    Techniques: Knockdown, CCK-8 Assay, Colony Assay, Migration, Expressing, Fluorescence, Imaging, Activity Assay, Flow Cytometry

    Knockdown of PEA15 Enhances the Anticancer Efficacy of DDP (A) Colony formation assays in control, DDP-treated, Si-PEA15, and DDP/Si-PEA15 combined treatment groups. (B) Migration and invasion assay images for the four treatment groups. (C) Fluorescence imaging of autophagic activity in the four treatment groups. (D and E) Expression levels of migration/invasion-related proteins and autophagy-related proteins in the four treatment groups. (F) Knockdown of PEA15 enhances the anticancer effect of DDP through the TNF signaling pathway. (G) The protein expression of ATG5 and LC3B was assessed in U2OS cells treated with the TNF-α antagonist (R-7050).

    Journal: iScience

    Article Title: PEA15 promotes osteosarcoma progression and cisplatin resistance by modulating autophagy through the FABP3-TNF signaling axis

    doi: 10.1016/j.isci.2025.113695

    Figure Lengend Snippet: Knockdown of PEA15 Enhances the Anticancer Efficacy of DDP (A) Colony formation assays in control, DDP-treated, Si-PEA15, and DDP/Si-PEA15 combined treatment groups. (B) Migration and invasion assay images for the four treatment groups. (C) Fluorescence imaging of autophagic activity in the four treatment groups. (D and E) Expression levels of migration/invasion-related proteins and autophagy-related proteins in the four treatment groups. (F) Knockdown of PEA15 enhances the anticancer effect of DDP through the TNF signaling pathway. (G) The protein expression of ATG5 and LC3B was assessed in U2OS cells treated with the TNF-α antagonist (R-7050).

    Article Snippet: The membrane was blocked with 5% non-fat milk in TBST for 2 h at room temperature, followed by incubation overnight at 4°C with primary antibodies targeting PEA15 (CST, #13091,1:1000), LC3B (CST, #3868,1:1000), ATG5 (CST,#9980,1:1000),Cleaved-Caspase3 (CST,#9664,1:1000),Cleaved-PARP (CST, #9541,1:1000), FABP3 (CST,#14780,1:1000), TNF-α (CST,#3707,1:1000), and GAPDH (CST, #5174,1:1000).

    Techniques: Knockdown, Control, Migration, Invasion Assay, Fluorescence, Imaging, Activity Assay, Expressing

    In Vivo Tumor Formation Assay (A) Images of solid tumors from control, DDP-treated, Sh-PEA15, and DDP/Sh-PEA15 combined treatment groups. (B and C) Statistical analysis of tumor volume and tumor weight in the four treatment groups ( n = 3). (D) Hematoxylin and eosin (H&E) staining of tumors from the four treatment groups ( n = 3). (E) Expression of Ki67 in tumors from the four treatment groups in the animal experiment. (F) Expression of LC3B in tumors from the four treatment groups in the animal experiment. The data are presented as mean ± SD. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

    Journal: iScience

    Article Title: PEA15 promotes osteosarcoma progression and cisplatin resistance by modulating autophagy through the FABP3-TNF signaling axis

    doi: 10.1016/j.isci.2025.113695

    Figure Lengend Snippet: In Vivo Tumor Formation Assay (A) Images of solid tumors from control, DDP-treated, Sh-PEA15, and DDP/Sh-PEA15 combined treatment groups. (B and C) Statistical analysis of tumor volume and tumor weight in the four treatment groups ( n = 3). (D) Hematoxylin and eosin (H&E) staining of tumors from the four treatment groups ( n = 3). (E) Expression of Ki67 in tumors from the four treatment groups in the animal experiment. (F) Expression of LC3B in tumors from the four treatment groups in the animal experiment. The data are presented as mean ± SD. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

    Article Snippet: The membrane was blocked with 5% non-fat milk in TBST for 2 h at room temperature, followed by incubation overnight at 4°C with primary antibodies targeting PEA15 (CST, #13091,1:1000), LC3B (CST, #3868,1:1000), ATG5 (CST,#9980,1:1000),Cleaved-Caspase3 (CST,#9664,1:1000),Cleaved-PARP (CST, #9541,1:1000), FABP3 (CST,#14780,1:1000), TNF-α (CST,#3707,1:1000), and GAPDH (CST, #5174,1:1000).

    Techniques: In Vivo, Tube Formation Assay, Control, Staining, Expressing

    PEA-15 co-traffics with Transferrin. ( A ) HeLa cells expressing GFP-tagged PEA-15 were incubated with Rhodamine-Transferrin for 2 min at 37 °C (Pulse). Non-internalized Transferrin was washed away before incubation at 37 °C in the presence of unlabeled transferrin (Chase). Immunofluorescence imaging shows the co-trafficking of GFP-PEA15 and chased Rhodamine-Transferrin to the perinuclear region. No trafficking was observed with GFP control. Scale bar in zoomed-out images = 25 µm. ( B ) Cells were processed as in Part A, but were also immunostained with antibodies against Rab5. Scale bar in zoomed-out images = 20 µm. Scale bar in zoomed-in images = 5 µm. ( C ) U87MG cells attached to fibronectin-coated glass coverslips for 30 min before co-immunostaining for endogenous PEA-15 and endogenous Rab5. Shown are maximum intensity projections from z-stacks acquired on a confocal microscope (XY) paired with orthogonal view (XZ). Arrows indicate perinuclear colocalization of PEA-15 with Rab5. ( D ) U87MG cells plated in normal growth conditions with the addition of primaquine were co-immunostained for endogenous PEA-15 and endogenous Rab5. Arrows indicate colocalization.

    Journal: Scientific Reports

    Article Title: Proteomics analysis identifies PEA-15 as an endosomal phosphoprotein that regulates α5β1 integrin endocytosis

    doi: 10.1038/s41598-021-99348-z

    Figure Lengend Snippet: PEA-15 co-traffics with Transferrin. ( A ) HeLa cells expressing GFP-tagged PEA-15 were incubated with Rhodamine-Transferrin for 2 min at 37 °C (Pulse). Non-internalized Transferrin was washed away before incubation at 37 °C in the presence of unlabeled transferrin (Chase). Immunofluorescence imaging shows the co-trafficking of GFP-PEA15 and chased Rhodamine-Transferrin to the perinuclear region. No trafficking was observed with GFP control. Scale bar in zoomed-out images = 25 µm. ( B ) Cells were processed as in Part A, but were also immunostained with antibodies against Rab5. Scale bar in zoomed-out images = 20 µm. Scale bar in zoomed-in images = 5 µm. ( C ) U87MG cells attached to fibronectin-coated glass coverslips for 30 min before co-immunostaining for endogenous PEA-15 and endogenous Rab5. Shown are maximum intensity projections from z-stacks acquired on a confocal microscope (XY) paired with orthogonal view (XZ). Arrows indicate perinuclear colocalization of PEA-15 with Rab5. ( D ) U87MG cells plated in normal growth conditions with the addition of primaquine were co-immunostained for endogenous PEA-15 and endogenous Rab5. Arrows indicate colocalization.

    Article Snippet: PEA-15-containing protein complexes were precipitated from whole cell lysates with anti-PEA15 antibodies (Pacific Immunology, Ramona, CA), or anti-HA antibodies (#51064-2-AP, Proteintech, Rosemont, IL) in the case of cells expressing HA-tagged PEA-15.

    Techniques: Expressing, Incubation, Immunofluorescence, Imaging, Control, Immunostaining, Microscopy

    PEA-15 regulates integrin trafficking. ( A ) Strip-resistant (internalized) biotinylated integrins were quantified by capture ELISA with antibodies against α5 integrin and expressed as a percentage of surface integrin absorbance signal. Shown is internalization after 5 min ( p = 0.2299, n = 2) and 15 min (* p < 0.05, n = 5) in the PEA-15 knock down cells compared to scramble control. Chased (recycled) biotinylated integrins were also quantified by capture ELISA. Shown is the percent α5 integrin recycled relative to pulse controls at 10 min (* p < 0.005, n = 4) and 20 min ( p = 0.5452, n = 3). Western blots show expression of α5 integrin, β1 integrin, Rab5 and Clathrin Heavy Chain, in the stable PEA-15 KD cells. ( B ) Surface biotinylated integrin α5β1 was quantified by capture ELISA (normalized absorbance to scramble control, p = 0.2439, n = 7). Degradation of surface α5 integrin was quantified over 6 h. Shown is the percent signal compared to time 0 (n = 3). ( C ) Integrin endocytosis was quantified in U87MG with two independent siRNAs (1 and 2). Shown is internalization normalized to non-target (NT) control (* p < 0.05, n = 3 for siRNA 1, n = 2 for siRNA 2). ( D ) Integrin endocytosis was measured using immunoprecipitation with antibodies against β1 integrin and detection with Western blotting with Streptavidin-HRP. Shown is a representative Western blot and internalization time course with points at 10 min ( p = 0.2425, n = 3) and 15 min (* p < 0.05, n = 3). ( E ) Surface α5β1 integrin was labeled with mouse anti-α5 antibodies in PEA-15 KD cells and scramble controls on ice. Cells were then returned to 37 °C in the presence of EGF to induce integrin trafficking, followed by fixation and immunostaining. HA-tag PEA-15 expressed in a subset of PEA-15 KD cells were co-stained with anti-HA-tag antibodies. Asterisks (*) mark transfected cells. Inset images show red-fluorescent HA-tag signal. Scale bar = 20 µm. ( F ) Integrin endocytosis was quantified in PEA-15 KD cells expressing HA-tagged PEA-15 (* p < 0.05, Scramble n = 3, PEA-15 shRNA n = 4, HA-PEA15 rescue n = 3, HA-S104A n = 3, HA-S116A n = 3).

    Journal: Scientific Reports

    Article Title: Proteomics analysis identifies PEA-15 as an endosomal phosphoprotein that regulates α5β1 integrin endocytosis

    doi: 10.1038/s41598-021-99348-z

    Figure Lengend Snippet: PEA-15 regulates integrin trafficking. ( A ) Strip-resistant (internalized) biotinylated integrins were quantified by capture ELISA with antibodies against α5 integrin and expressed as a percentage of surface integrin absorbance signal. Shown is internalization after 5 min ( p = 0.2299, n = 2) and 15 min (* p < 0.05, n = 5) in the PEA-15 knock down cells compared to scramble control. Chased (recycled) biotinylated integrins were also quantified by capture ELISA. Shown is the percent α5 integrin recycled relative to pulse controls at 10 min (* p < 0.005, n = 4) and 20 min ( p = 0.5452, n = 3). Western blots show expression of α5 integrin, β1 integrin, Rab5 and Clathrin Heavy Chain, in the stable PEA-15 KD cells. ( B ) Surface biotinylated integrin α5β1 was quantified by capture ELISA (normalized absorbance to scramble control, p = 0.2439, n = 7). Degradation of surface α5 integrin was quantified over 6 h. Shown is the percent signal compared to time 0 (n = 3). ( C ) Integrin endocytosis was quantified in U87MG with two independent siRNAs (1 and 2). Shown is internalization normalized to non-target (NT) control (* p < 0.05, n = 3 for siRNA 1, n = 2 for siRNA 2). ( D ) Integrin endocytosis was measured using immunoprecipitation with antibodies against β1 integrin and detection with Western blotting with Streptavidin-HRP. Shown is a representative Western blot and internalization time course with points at 10 min ( p = 0.2425, n = 3) and 15 min (* p < 0.05, n = 3). ( E ) Surface α5β1 integrin was labeled with mouse anti-α5 antibodies in PEA-15 KD cells and scramble controls on ice. Cells were then returned to 37 °C in the presence of EGF to induce integrin trafficking, followed by fixation and immunostaining. HA-tag PEA-15 expressed in a subset of PEA-15 KD cells were co-stained with anti-HA-tag antibodies. Asterisks (*) mark transfected cells. Inset images show red-fluorescent HA-tag signal. Scale bar = 20 µm. ( F ) Integrin endocytosis was quantified in PEA-15 KD cells expressing HA-tagged PEA-15 (* p < 0.05, Scramble n = 3, PEA-15 shRNA n = 4, HA-PEA15 rescue n = 3, HA-S104A n = 3, HA-S116A n = 3).

    Article Snippet: PEA-15-containing protein complexes were precipitated from whole cell lysates with anti-PEA15 antibodies (Pacific Immunology, Ramona, CA), or anti-HA antibodies (#51064-2-AP, Proteintech, Rosemont, IL) in the case of cells expressing HA-tagged PEA-15.

    Techniques: Stripping Membranes, Enzyme-linked Immunosorbent Assay, Knockdown, Control, Western Blot, Expressing, Immunoprecipitation, Labeling, Immunostaining, Staining, Transfection, shRNA

    mPEA15 expressed ubiquitously in transgenic pigs. a Representative western blot image showing mPEA15 expression in muscle tissues of the wild type and transgenic pigs. The tissue lysate was immunoblotted with anti-mPEA15 and anti-FLAG antibodies as mentioned in materials and methods. WT: Wild-type pigs (n = 3) TG: Transgenic pigs (n = 5). b Graph showing the mPEA15 fold difference between wild type and transgenic pigs. Results represent means SEM from three separate experiments. *P < 0.001 compared to WT. c Representative blots showing PEA15 expression. Tissue sections obtained from muscle, liver, and pancreas were stained with anti-PEA15 and anti-FLAG antibodies. Scale bar is 200 µm

    Journal: 3 Biotech

    Article Title: Validation of mouse phosphoprotein enriched in astrocyte 15 (mPEA15) expressing transgenic pig as a potential model in diabetes translational research

    doi: 10.1007/s13205-019-2021-0

    Figure Lengend Snippet: mPEA15 expressed ubiquitously in transgenic pigs. a Representative western blot image showing mPEA15 expression in muscle tissues of the wild type and transgenic pigs. The tissue lysate was immunoblotted with anti-mPEA15 and anti-FLAG antibodies as mentioned in materials and methods. WT: Wild-type pigs (n = 3) TG: Transgenic pigs (n = 5). b Graph showing the mPEA15 fold difference between wild type and transgenic pigs. Results represent means SEM from three separate experiments. *P < 0.001 compared to WT. c Representative blots showing PEA15 expression. Tissue sections obtained from muscle, liver, and pancreas were stained with anti-PEA15 and anti-FLAG antibodies. Scale bar is 200 µm

    Article Snippet: Briefly, after rehydration, the slides were incubated for 90 min in blocking buffer supplemented with 5% non-fat dry milk and 0.05% Tween-20 at room temperature and probed with a primary antibody against PEA15 (1:100, Cell Signaling Biotech.

    Techniques: Transgenic Assay, Western Blot, Expressing, Staining