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rabbit polyclonal anti-anxa1  (Thermo Fisher)


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    Thermo Fisher rabbit polyclonal anti-anxa1
    Rabbit Polyclonal Anti Anxa1, 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
    https://www.bioz.com/product/rabbit+polyclonal+anti-anxa1/accell+sirnas/pm40100418-112-7-11
    Average 90 stars, based on 1 article reviews
    rabbit polyclonal anti-anxa1 - by Bioz Stars, 2026-10
    90/100 stars

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    Related Articles

    Serial Time-encoded Amplified Microscopy:

    Article Title: Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature.
    Article Snippet: Human polymorphonuclear leukocytes adhesion to endothelial cells during the early stage of inflammation leads to cell surface externalization of Annexin A1 (AnxA1), an effector of endogenous anti-inflammation.. The antiadhesive properties of AnxA1 become operative to finely tune polymorphonuclear leukocytes transmigration to the site of inflammation.. Membrane bound proteinase 3 (PR3) plays a key role in this microenvironment by cleaving the N terminus bioactive domain of AnxA1.

    Article Title: Pharmacological treatment with annexin A1-derived peptide protects against cisplatin-induced hearing loss.
    Article Snippet: Sections were washed with distilled water, and antigen retrieval was performed with citrate buffer, pH 6.0, in a steam cooker at 96 ◦C, for 30 min, followed by incubation in 4% bovine serum albumin (BSA) in phosphate-buffered saline (PBS), at room temperature for 1 h. Sections were incubated overnight at 4 ◦C with rabbit polyclonal anti-AnxA1 (Invitrogen - Thermo Fisher Scientific, Waltham, MA, USA), anti-Fpr2, anti-pERK (Santa Cruz Biotechnology, CA, USA), and anticleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA) antibodies diluted 1:500, 1:200, 1:200, and 1:50, respectively, in 1% BSA in PBS.

    Article Title: Regulatory role of annexin A1 in NLRP3 inflammasome activation in atopic dermatitis: insights from keratinocytes in human and murine studies.
    Article Snippet: Despite the well-documented regulatory role of annexin A1 (ANXA1) in numerous stages of the inflammatory response, its involvement in regulating the NLRP3 inflammasome in the context of allergic responses has not been extensively investigated to date.. This study evaluated the expression patterns of the ANXA1 and NLRP3 proteins in human skin samples obtained from patients with atopic dermatitis (AD) and in mice with ovalbumin (OVA)-induced experimental AD.. Furthermore, the in vitro effect of the ANXA1 mimetic peptide Ac2-26 on IL-4-stimulated human keratinocytes was evaluated.

    Article Title: Annexin 1 Cleavage in Activated Neutrophils
    Article Snippet: The primary antibodies used were rabbit polyclonal anti-AnxA1 (Zymed Laboratories Inc., Cambridge, UK), mouse monoclonal anti-c-Myc (Santa Cruz), and mouse monoclonal anti-FLAG M2 (Sigma).

    Incubation:

    Article Title: Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature.
    Article Snippet: Human polymorphonuclear leukocytes adhesion to endothelial cells during the early stage of inflammation leads to cell surface externalization of Annexin A1 (AnxA1), an effector of endogenous anti-inflammation.. The antiadhesive properties of AnxA1 become operative to finely tune polymorphonuclear leukocytes transmigration to the site of inflammation.. Membrane bound proteinase 3 (PR3) plays a key role in this microenvironment by cleaving the N terminus bioactive domain of AnxA1.

    Article Title: Pharmacological treatment with annexin A1-derived peptide protects against cisplatin-induced hearing loss.
    Article Snippet: Sections were washed with distilled water, and antigen retrieval was performed with citrate buffer, pH 6.0, in a steam cooker at 96 ◦C, for 30 min, followed by incubation in 4% bovine serum albumin (BSA) in phosphate-buffered saline (PBS), at room temperature for 1 h. Sections were incubated overnight at 4 ◦C with rabbit polyclonal anti-AnxA1 (Invitrogen - Thermo Fisher Scientific, Waltham, MA, USA), anti-Fpr2, anti-pERK (Santa Cruz Biotechnology, CA, USA), and anticleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA) antibodies diluted 1:500, 1:200, 1:200, and 1:50, respectively, in 1% BSA in PBS.

    Article Title: Regulatory role of annexin A1 in NLRP3 inflammasome activation in atopic dermatitis: insights from keratinocytes in human and murine studies.
    Article Snippet: Despite the well-documented regulatory role of annexin A1 (ANXA1) in numerous stages of the inflammatory response, its involvement in regulating the NLRP3 inflammasome in the context of allergic responses has not been extensively investigated to date.. This study evaluated the expression patterns of the ANXA1 and NLRP3 proteins in human skin samples obtained from patients with atopic dermatitis (AD) and in mice with ovalbumin (OVA)-induced experimental AD.. Furthermore, the in vitro effect of the ANXA1 mimetic peptide Ac2-26 on IL-4-stimulated human keratinocytes was evaluated.

    Article Title: Annexin 1 Cleavage in Activated Neutrophils
    Article Snippet: The primary antibodies used were rabbit polyclonal anti-AnxA1 (Zymed Laboratories Inc., Cambridge, UK), mouse monoclonal anti-c-Myc (Santa Cruz), and mouse monoclonal anti-FLAG M2 (Sigma).

    Saline:

    Article Title: Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature.
    Article Snippet: Human polymorphonuclear leukocytes adhesion to endothelial cells during the early stage of inflammation leads to cell surface externalization of Annexin A1 (AnxA1), an effector of endogenous anti-inflammation.. The antiadhesive properties of AnxA1 become operative to finely tune polymorphonuclear leukocytes transmigration to the site of inflammation.. Membrane bound proteinase 3 (PR3) plays a key role in this microenvironment by cleaving the N terminus bioactive domain of AnxA1.

    Article Title: Pharmacological treatment with annexin A1-derived peptide protects against cisplatin-induced hearing loss.
    Article Snippet: Sections were washed with distilled water, and antigen retrieval was performed with citrate buffer, pH 6.0, in a steam cooker at 96 ◦C, for 30 min, followed by incubation in 4% bovine serum albumin (BSA) in phosphate-buffered saline (PBS), at room temperature for 1 h. Sections were incubated overnight at 4 ◦C with rabbit polyclonal anti-AnxA1 (Invitrogen - Thermo Fisher Scientific, Waltham, MA, USA), anti-Fpr2, anti-pERK (Santa Cruz Biotechnology, CA, USA), and anticleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA) antibodies diluted 1:500, 1:200, 1:200, and 1:50, respectively, in 1% BSA in PBS.

    Article Title: Regulatory role of annexin A1 in NLRP3 inflammasome activation in atopic dermatitis: insights from keratinocytes in human and murine studies.
    Article Snippet: Despite the well-documented regulatory role of annexin A1 (ANXA1) in numerous stages of the inflammatory response, its involvement in regulating the NLRP3 inflammasome in the context of allergic responses has not been extensively investigated to date.. This study evaluated the expression patterns of the ANXA1 and NLRP3 proteins in human skin samples obtained from patients with atopic dermatitis (AD) and in mice with ovalbumin (OVA)-induced experimental AD.. Furthermore, the in vitro effect of the ANXA1 mimetic peptide Ac2-26 on IL-4-stimulated human keratinocytes was evaluated.

    Article Title: Annexin 1 Cleavage in Activated Neutrophils
    Article Snippet: The primary antibodies used were rabbit polyclonal anti-AnxA1 (Zymed Laboratories Inc., Cambridge, UK), mouse monoclonal anti-c-Myc (Santa Cruz), and mouse monoclonal anti-FLAG M2 (Sigma).

    Produced:

    Article Title: Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature.
    Article Snippet: Human polymorphonuclear leukocytes adhesion to endothelial cells during the early stage of inflammation leads to cell surface externalization of Annexin A1 (AnxA1), an effector of endogenous anti-inflammation.. The antiadhesive properties of AnxA1 become operative to finely tune polymorphonuclear leukocytes transmigration to the site of inflammation.. Membrane bound proteinase 3 (PR3) plays a key role in this microenvironment by cleaving the N terminus bioactive domain of AnxA1.

    Article Title: Pharmacological treatment with annexin A1-derived peptide protects against cisplatin-induced hearing loss.
    Article Snippet: Sections were washed with distilled water, and antigen retrieval was performed with citrate buffer, pH 6.0, in a steam cooker at 96 ◦C, for 30 min, followed by incubation in 4% bovine serum albumin (BSA) in phosphate-buffered saline (PBS), at room temperature for 1 h. Sections were incubated overnight at 4 ◦C with rabbit polyclonal anti-AnxA1 (Invitrogen - Thermo Fisher Scientific, Waltham, MA, USA), anti-Fpr2, anti-pERK (Santa Cruz Biotechnology, CA, USA), and anticleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA) antibodies diluted 1:500, 1:200, 1:200, and 1:50, respectively, in 1% BSA in PBS.

    Article Title: Regulatory role of annexin A1 in NLRP3 inflammasome activation in atopic dermatitis: insights from keratinocytes in human and murine studies.
    Article Snippet: Despite the well-documented regulatory role of annexin A1 (ANXA1) in numerous stages of the inflammatory response, its involvement in regulating the NLRP3 inflammasome in the context of allergic responses has not been extensively investigated to date.. This study evaluated the expression patterns of the ANXA1 and NLRP3 proteins in human skin samples obtained from patients with atopic dermatitis (AD) and in mice with ovalbumin (OVA)-induced experimental AD.. Furthermore, the in vitro effect of the ANXA1 mimetic peptide Ac2-26 on IL-4-stimulated human keratinocytes was evaluated.

    Article Title: Annexin 1 Cleavage in Activated Neutrophils
    Article Snippet: The primary antibodies used were rabbit polyclonal anti-AnxA1 (Zymed Laboratories Inc., Cambridge, UK), mouse monoclonal anti-c-Myc (Santa Cruz), and mouse monoclonal anti-FLAG M2 (Sigma).

    MTT Assay:

    Article Title: Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature.
    Article Snippet: Human polymorphonuclear leukocytes adhesion to endothelial cells during the early stage of inflammation leads to cell surface externalization of Annexin A1 (AnxA1), an effector of endogenous anti-inflammation.. The antiadhesive properties of AnxA1 become operative to finely tune polymorphonuclear leukocytes transmigration to the site of inflammation.. Membrane bound proteinase 3 (PR3) plays a key role in this microenvironment by cleaving the N terminus bioactive domain of AnxA1.

    Article Title: Pharmacological treatment with annexin A1-derived peptide protects against cisplatin-induced hearing loss.
    Article Snippet: Sections were washed with distilled water, and antigen retrieval was performed with citrate buffer, pH 6.0, in a steam cooker at 96 ◦C, for 30 min, followed by incubation in 4% bovine serum albumin (BSA) in phosphate-buffered saline (PBS), at room temperature for 1 h. Sections were incubated overnight at 4 ◦C with rabbit polyclonal anti-AnxA1 (Invitrogen - Thermo Fisher Scientific, Waltham, MA, USA), anti-Fpr2, anti-pERK (Santa Cruz Biotechnology, CA, USA), and anticleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA) antibodies diluted 1:500, 1:200, 1:200, and 1:50, respectively, in 1% BSA in PBS.

    Article Title: Regulatory role of annexin A1 in NLRP3 inflammasome activation in atopic dermatitis: insights from keratinocytes in human and murine studies.
    Article Snippet: Despite the well-documented regulatory role of annexin A1 (ANXA1) in numerous stages of the inflammatory response, its involvement in regulating the NLRP3 inflammasome in the context of allergic responses has not been extensively investigated to date.. This study evaluated the expression patterns of the ANXA1 and NLRP3 proteins in human skin samples obtained from patients with atopic dermatitis (AD) and in mice with ovalbumin (OVA)-induced experimental AD.. Furthermore, the in vitro effect of the ANXA1 mimetic peptide Ac2-26 on IL-4-stimulated human keratinocytes was evaluated.

    Article Title: Annexin 1 Cleavage in Activated Neutrophils
    Article Snippet: The primary antibodies used were rabbit polyclonal anti-AnxA1 (Zymed Laboratories Inc., Cambridge, UK), mouse monoclonal anti-c-Myc (Santa Cruz), and mouse monoclonal anti-FLAG M2 (Sigma).

    Expressing:

    Article Title: Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature.
    Article Snippet: Human polymorphonuclear leukocytes adhesion to endothelial cells during the early stage of inflammation leads to cell surface externalization of Annexin A1 (AnxA1), an effector of endogenous anti-inflammation.. The antiadhesive properties of AnxA1 become operative to finely tune polymorphonuclear leukocytes transmigration to the site of inflammation.. Membrane bound proteinase 3 (PR3) plays a key role in this microenvironment by cleaving the N terminus bioactive domain of AnxA1.

    Article Title: Pharmacological treatment with annexin A1-derived peptide protects against cisplatin-induced hearing loss.
    Article Snippet: Sections were washed with distilled water, and antigen retrieval was performed with citrate buffer, pH 6.0, in a steam cooker at 96 ◦C, for 30 min, followed by incubation in 4% bovine serum albumin (BSA) in phosphate-buffered saline (PBS), at room temperature for 1 h. Sections were incubated overnight at 4 ◦C with rabbit polyclonal anti-AnxA1 (Invitrogen - Thermo Fisher Scientific, Waltham, MA, USA), anti-Fpr2, anti-pERK (Santa Cruz Biotechnology, CA, USA), and anticleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA) antibodies diluted 1:500, 1:200, 1:200, and 1:50, respectively, in 1% BSA in PBS.

    Article Title: Regulatory role of annexin A1 in NLRP3 inflammasome activation in atopic dermatitis: insights from keratinocytes in human and murine studies.
    Article Snippet: Despite the well-documented regulatory role of annexin A1 (ANXA1) in numerous stages of the inflammatory response, its involvement in regulating the NLRP3 inflammasome in the context of allergic responses has not been extensively investigated to date.. This study evaluated the expression patterns of the ANXA1 and NLRP3 proteins in human skin samples obtained from patients with atopic dermatitis (AD) and in mice with ovalbumin (OVA)-induced experimental AD.. Furthermore, the in vitro effect of the ANXA1 mimetic peptide Ac2-26 on IL-4-stimulated human keratinocytes was evaluated.

    Article Title: Annexin 1 Cleavage in Activated Neutrophils
    Article Snippet: The primary antibodies used were rabbit polyclonal anti-AnxA1 (Zymed Laboratories Inc., Cambridge, UK), mouse monoclonal anti-c-Myc (Santa Cruz), and mouse monoclonal anti-FLAG M2 (Sigma).

    Negative Control:

    Article Title: Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature.
    Article Snippet: Human polymorphonuclear leukocytes adhesion to endothelial cells during the early stage of inflammation leads to cell surface externalization of Annexin A1 (AnxA1), an effector of endogenous anti-inflammation.. The antiadhesive properties of AnxA1 become operative to finely tune polymorphonuclear leukocytes transmigration to the site of inflammation.. Membrane bound proteinase 3 (PR3) plays a key role in this microenvironment by cleaving the N terminus bioactive domain of AnxA1.

    Article Title: Pharmacological treatment with annexin A1-derived peptide protects against cisplatin-induced hearing loss.
    Article Snippet: Sections were washed with distilled water, and antigen retrieval was performed with citrate buffer, pH 6.0, in a steam cooker at 96 ◦C, for 30 min, followed by incubation in 4% bovine serum albumin (BSA) in phosphate-buffered saline (PBS), at room temperature for 1 h. Sections were incubated overnight at 4 ◦C with rabbit polyclonal anti-AnxA1 (Invitrogen - Thermo Fisher Scientific, Waltham, MA, USA), anti-Fpr2, anti-pERK (Santa Cruz Biotechnology, CA, USA), and anticleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA) antibodies diluted 1:500, 1:200, 1:200, and 1:50, respectively, in 1% BSA in PBS.

    Article Title: Regulatory role of annexin A1 in NLRP3 inflammasome activation in atopic dermatitis: insights from keratinocytes in human and murine studies.
    Article Snippet: Despite the well-documented regulatory role of annexin A1 (ANXA1) in numerous stages of the inflammatory response, its involvement in regulating the NLRP3 inflammasome in the context of allergic responses has not been extensively investigated to date.. This study evaluated the expression patterns of the ANXA1 and NLRP3 proteins in human skin samples obtained from patients with atopic dermatitis (AD) and in mice with ovalbumin (OVA)-induced experimental AD.. Furthermore, the in vitro effect of the ANXA1 mimetic peptide Ac2-26 on IL-4-stimulated human keratinocytes was evaluated.

    Article Title: Annexin 1 Cleavage in Activated Neutrophils
    Article Snippet: The primary antibodies used were rabbit polyclonal anti-AnxA1 (Zymed Laboratories Inc., Cambridge, UK), mouse monoclonal anti-c-Myc (Santa Cruz), and mouse monoclonal anti-FLAG M2 (Sigma).



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    Cell Signaling Technology Inc rabbit polyclonal anti-anxa1
    <t>ANXA1</t> as an ATE1-mediated determinant cytokine inducing pro-apoptotic, anti-inflammatory, and pro-proliferative effects on recipient cells (A) A graph representing delta Δ fold change and Δ fold change of iBAQ. (B) A table regarding iBAQ level and fold change value of ANXA1 in secreted exosomes from A549 treated with tBHP (250 μM, 6 h) and RNA interference of ATE1 . (C) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 in A549 cells with 250 μM or 500 μM tBHP treatment (6 h). (D) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 and Tubulin in A549 cells with RNA interference of ATE1 and 250 μM tBHP treatment (6 h). (E) Immunoblotting analysis of A549 cells under ANXA1 knockdown and treated with tBHP (250 μM, 6 h). (F) Immunoblotting analysis of A549 cells treated with exosomes from E (24 h). (G) Cellular viability of recipient A549 cells treated with exosomes originated from donor cells under ANXA1 knockdown and treated with tBHP (500 μM, 6 h) (24h). Results are presented as OD values (absorbance at 450 nm). (H) Immunostaining analysis of γH2AX in A549 cells with RNA interference of ANXA1 and treatment of tBHP (500 μM, 6 h). Scale bar, 10 μm. (I) Quantification of H ( n = 150). (J) Relative mRNA level of IL-6 in A549 cells treated with exosomes originated from cells under ANXA1 knockdown and tBHP (500 μM, 6 h) treatment compared with control. (K) Relative mRNA level of TNFα in A549 cells treated with exosome originated from cells under ANXA1 knockdown and tBHP (500 μM, 6 h) treatment compared with control. Error bars represent SEM ( n = 3 replicates). ∗ p < 0.05, ∗∗ p < 0.01 using the paired t-test. See also .
    Rabbit Polyclonal Anti Anxa1, supplied by Cell Signaling Technology Inc, 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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    Cell Signaling Technology Inc rabbit polyclonal anti anxa1
    <t>ANXA1</t> as an ATE1-mediated determinant cytokine inducing pro-apoptotic, anti-inflammatory, and pro-proliferative effects on recipient cells (A) A graph representing delta Δ fold change and Δ fold change of iBAQ. (B) A table regarding iBAQ level and fold change value of ANXA1 in secreted exosomes from A549 treated with tBHP (250 μM, 6 h) and RNA interference of ATE1 . (C) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 in A549 cells with 250 μM or 500 μM tBHP treatment (6 h). (D) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 and Tubulin in A549 cells with RNA interference of ATE1 and 250 μM tBHP treatment (6 h). (E) Immunoblotting analysis of A549 cells under ANXA1 knockdown and treated with tBHP (250 μM, 6 h). (F) Immunoblotting analysis of A549 cells treated with exosomes from E (24 h). (G) Cellular viability of recipient A549 cells treated with exosomes originated from donor cells under ANXA1 knockdown and treated with tBHP (500 μM, 6 h) (24h). Results are presented as OD values (absorbance at 450 nm). (H) Immunostaining analysis of γH2AX in A549 cells with RNA interference of ANXA1 and treatment of tBHP (500 μM, 6 h). Scale bar, 10 μm. (I) Quantification of H ( n = 150). (J) Relative mRNA level of IL-6 in A549 cells treated with exosomes originated from cells under ANXA1 knockdown and tBHP (500 μM, 6 h) treatment compared with control. (K) Relative mRNA level of TNFα in A549 cells treated with exosome originated from cells under ANXA1 knockdown and tBHP (500 μM, 6 h) treatment compared with control. Error bars represent SEM ( n = 3 replicates). ∗ p < 0.05, ∗∗ p < 0.01 using the paired t-test. See also .
    Rabbit Polyclonal Anti Anxa1, supplied by Cell Signaling Technology Inc, 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/rabbit+polyclonal+anti-anxa1/Annexin+A1+Antibody/pmc10382581-308-8-18
    Average 93 stars, based on 1 article reviews
    rabbit polyclonal anti anxa1 - by Bioz Stars, 2026-10
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    ANXA1 as an ATE1-mediated determinant cytokine inducing pro-apoptotic, anti-inflammatory, and pro-proliferative effects on recipient cells (A) A graph representing delta Δ fold change and Δ fold change of iBAQ. (B) A table regarding iBAQ level and fold change value of ANXA1 in secreted exosomes from A549 treated with tBHP (250 μM, 6 h) and RNA interference of ATE1 . (C) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 in A549 cells with 250 μM or 500 μM tBHP treatment (6 h). (D) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 and Tubulin in A549 cells with RNA interference of ATE1 and 250 μM tBHP treatment (6 h). (E) Immunoblotting analysis of A549 cells under ANXA1 knockdown and treated with tBHP (250 μM, 6 h). (F) Immunoblotting analysis of A549 cells treated with exosomes from E (24 h). (G) Cellular viability of recipient A549 cells treated with exosomes originated from donor cells under ANXA1 knockdown and treated with tBHP (500 μM, 6 h) (24h). Results are presented as OD values (absorbance at 450 nm). (H) Immunostaining analysis of γH2AX in A549 cells with RNA interference of ANXA1 and treatment of tBHP (500 μM, 6 h). Scale bar, 10 μm. (I) Quantification of H ( n = 150). (J) Relative mRNA level of IL-6 in A549 cells treated with exosomes originated from cells under ANXA1 knockdown and tBHP (500 μM, 6 h) treatment compared with control. (K) Relative mRNA level of TNFα in A549 cells treated with exosome originated from cells under ANXA1 knockdown and tBHP (500 μM, 6 h) treatment compared with control. Error bars represent SEM ( n = 3 replicates). ∗ p < 0.05, ∗∗ p < 0.01 using the paired t-test. See also .

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    Figure Lengend Snippet: ANXA1 as an ATE1-mediated determinant cytokine inducing pro-apoptotic, anti-inflammatory, and pro-proliferative effects on recipient cells (A) A graph representing delta Δ fold change and Δ fold change of iBAQ. (B) A table regarding iBAQ level and fold change value of ANXA1 in secreted exosomes from A549 treated with tBHP (250 μM, 6 h) and RNA interference of ATE1 . (C) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 in A549 cells with 250 μM or 500 μM tBHP treatment (6 h). (D) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 and Tubulin in A549 cells with RNA interference of ATE1 and 250 μM tBHP treatment (6 h). (E) Immunoblotting analysis of A549 cells under ANXA1 knockdown and treated with tBHP (250 μM, 6 h). (F) Immunoblotting analysis of A549 cells treated with exosomes from E (24 h). (G) Cellular viability of recipient A549 cells treated with exosomes originated from donor cells under ANXA1 knockdown and treated with tBHP (500 μM, 6 h) (24h). Results are presented as OD values (absorbance at 450 nm). (H) Immunostaining analysis of γH2AX in A549 cells with RNA interference of ANXA1 and treatment of tBHP (500 μM, 6 h). Scale bar, 10 μm. (I) Quantification of H ( n = 150). (J) Relative mRNA level of IL-6 in A549 cells treated with exosomes originated from cells under ANXA1 knockdown and tBHP (500 μM, 6 h) treatment compared with control. (K) Relative mRNA level of TNFα in A549 cells treated with exosome originated from cells under ANXA1 knockdown and tBHP (500 μM, 6 h) treatment compared with control. Error bars represent SEM ( n = 3 replicates). ∗ p < 0.05, ∗∗ p < 0.01 using the paired t-test. See also .

    Article Snippet: Rabbit polyclonal Anti-ANXA1 , Bethyl , Cat#A305-235A; RRID: AB_2631627.

    Techniques: Western Blot, Knockdown, Immunostaining, Control

    ATE1-dependent interaction of VPS36 and VPS22 facilitates invagination of ANXA1 + ILVs (A) Co-immunoprecipitation (Co-IP) analysis of the interaction between endogenous ANXA1 and VPS22 in A549 cells followed by tBHP (250 μM, 6 h) treatment. (B) Denaturation-IP analysis of the interaction between endogenous ANXA1 in A549 cells transfected with Ub −HA under ATE1 knockdown followed by tBHP (250 μM, 6 h) treatment. (C) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 in A549 cells with RNA interference of UBR1 and UBR2 , followed by tBHP treatment (250 μM, 6 h). (D) Co-IP analysis of the interaction between VPS36 and VPS22 in A549 cells transfected with VPS22 −MYC with RNA interference of ATE1 followed by tBHP (250 μM, 6 h) treatment. (E) Co-localization immunostaining analysis of VPS22 with VPS36 in HeLa cells with RNA interference of RILP and treatment of tBHP (250 μM, 6 h). Scale bar, 10 μm. (F) Quantification of E ( n = 50). (G) Transmission electron microscopy (TEM) image of WT and ATE1 −/− MEF cells with tBHP treatment (250 μM, 6 h). (H) Quantification of G ( n = 30). Error bars represent SEM ( n = 3 replicates). ∗ p < 0.05, ∗∗ p < 0.01 using the paired t-test. See also .

    Journal:

    Article Title:

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    Figure Lengend Snippet: ATE1-dependent interaction of VPS36 and VPS22 facilitates invagination of ANXA1 + ILVs (A) Co-immunoprecipitation (Co-IP) analysis of the interaction between endogenous ANXA1 and VPS22 in A549 cells followed by tBHP (250 μM, 6 h) treatment. (B) Denaturation-IP analysis of the interaction between endogenous ANXA1 in A549 cells transfected with Ub −HA under ATE1 knockdown followed by tBHP (250 μM, 6 h) treatment. (C) Immunoblotting analysis of cell lysate and exosome fraction ANXA1 in A549 cells with RNA interference of UBR1 and UBR2 , followed by tBHP treatment (250 μM, 6 h). (D) Co-IP analysis of the interaction between VPS36 and VPS22 in A549 cells transfected with VPS22 −MYC with RNA interference of ATE1 followed by tBHP (250 μM, 6 h) treatment. (E) Co-localization immunostaining analysis of VPS22 with VPS36 in HeLa cells with RNA interference of RILP and treatment of tBHP (250 μM, 6 h). Scale bar, 10 μm. (F) Quantification of E ( n = 50). (G) Transmission electron microscopy (TEM) image of WT and ATE1 −/− MEF cells with tBHP treatment (250 μM, 6 h). (H) Quantification of G ( n = 30). Error bars represent SEM ( n = 3 replicates). ∗ p < 0.05, ∗∗ p < 0.01 using the paired t-test. See also .

    Article Snippet: Rabbit polyclonal Anti-ANXA1 , Bethyl , Cat#A305-235A; RRID: AB_2631627.

    Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection, Knockdown, Western Blot, Immunostaining, Transmission Assay, Electron Microscopy

    ATE1 is required for RILP cleavage and subsequent secretion of ANXA1 under oxidative stress (A) Immunoblotting analysis of cell lysate and exosome fraction ANXA1, RILP, and GAPDH in A549 cells with RNA interference of RILP and 250 μM tBHP treatment (6 h). (B) Immunoblotting analysis of A549 cells transfected with RILP −MYC/FLAG with tBHP (250 μM, 6 h), CoCl 2 (250 μM, 6 h), CoCl 2 (250 μM, 24 h), or poly dA:dT (1 μg/μL, 24 h). (C) Immunoblotting analysis of A549 cells transfected with RILP −MYC/FLAG with RNA interference of ATE1 and 250 μM tBHP treatment (6 h). (D) Immunoblotting analysis of cleaved C-term RILP in A549 cells transfected with RILP −MYC/FLAG , Caspase 1 −MYC , or both. (E) Immunoblotting analysis of A549 cells under RILP knockdown and treated with tBHP (250 μM, 6 h). (F) Relative mRNA level of Caspase-1 in A549 cells treated with exosome originated from cells under ATE1 knockdown and tBHP (250 μM, 6 h) treatment compared with control. (G) Co-localization immunostaining analysis of VPS36 and VPS22 in WT and ATE1 −/− HeLa cells transfected with VPS36 −GFP and VPS22 −MYC with treatment of tBHP (250 μM, 6 h). Scale bar, 10 μm. (H) Quantification of G ( n = 50). (I) Immunoblotting analysis of A549 cells under RILP knockdown and treated with tBHP (250 μM, 6 h). (J) Immunoblotting analysis of A549 cells treated with exosomes from I (24 h). (K) Relative mRNA level of RILP in A549 cells under RILP knockdown and tBHP (250 μM, 6 h) treatment compared with control. (L) Immunostaining analysis of γH2AX in A549 cells with RNA interference of ANXA1 and treatment of tBHP (500 μM, 6 h). Scale bar, 10 μm. (M) Quantification of L ( n = 150). (N) Cellular viability of recipient A549 cells with exosome originated from donor cells under RILP knockdown and treated with tBHP (500 μM, 6 h) (24 h). Results are presented as OD values (absorbance at 450 nm). Error bars represent SEM ( n = 3 replicates). ∗ p < 0.05, ∗∗ p < 0.01 using the paired t-test. See also .

    Journal:

    Article Title:

    doi:

    Figure Lengend Snippet: ATE1 is required for RILP cleavage and subsequent secretion of ANXA1 under oxidative stress (A) Immunoblotting analysis of cell lysate and exosome fraction ANXA1, RILP, and GAPDH in A549 cells with RNA interference of RILP and 250 μM tBHP treatment (6 h). (B) Immunoblotting analysis of A549 cells transfected with RILP −MYC/FLAG with tBHP (250 μM, 6 h), CoCl 2 (250 μM, 6 h), CoCl 2 (250 μM, 24 h), or poly dA:dT (1 μg/μL, 24 h). (C) Immunoblotting analysis of A549 cells transfected with RILP −MYC/FLAG with RNA interference of ATE1 and 250 μM tBHP treatment (6 h). (D) Immunoblotting analysis of cleaved C-term RILP in A549 cells transfected with RILP −MYC/FLAG , Caspase 1 −MYC , or both. (E) Immunoblotting analysis of A549 cells under RILP knockdown and treated with tBHP (250 μM, 6 h). (F) Relative mRNA level of Caspase-1 in A549 cells treated with exosome originated from cells under ATE1 knockdown and tBHP (250 μM, 6 h) treatment compared with control. (G) Co-localization immunostaining analysis of VPS36 and VPS22 in WT and ATE1 −/− HeLa cells transfected with VPS36 −GFP and VPS22 −MYC with treatment of tBHP (250 μM, 6 h). Scale bar, 10 μm. (H) Quantification of G ( n = 50). (I) Immunoblotting analysis of A549 cells under RILP knockdown and treated with tBHP (250 μM, 6 h). (J) Immunoblotting analysis of A549 cells treated with exosomes from I (24 h). (K) Relative mRNA level of RILP in A549 cells under RILP knockdown and tBHP (250 μM, 6 h) treatment compared with control. (L) Immunostaining analysis of γH2AX in A549 cells with RNA interference of ANXA1 and treatment of tBHP (500 μM, 6 h). Scale bar, 10 μm. (M) Quantification of L ( n = 150). (N) Cellular viability of recipient A549 cells with exosome originated from donor cells under RILP knockdown and treated with tBHP (500 μM, 6 h) (24 h). Results are presented as OD values (absorbance at 450 nm). Error bars represent SEM ( n = 3 replicates). ∗ p < 0.05, ∗∗ p < 0.01 using the paired t-test. See also .

    Article Snippet: Rabbit polyclonal Anti-ANXA1 , Bethyl , Cat#A305-235A; RRID: AB_2631627.

    Techniques: Western Blot, Transfection, Knockdown, Control, Immunostaining