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5- (3-bromophenyl)-7-[6-(4-morpholinyl)-3-pyrido[2,3-d]byrimidin-4amine hydrochloride abt 702  (Tocris)


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    Tocris 5- (3-bromophenyl)-7-[6-(4-morpholinyl)-3-pyrido[2,3-d]byrimidin-4amine hydrochloride abt 702
    5 (3 Bromophenyl) 7 [6 (4 Morpholinyl) 3 Pyrido[2,3 D]Byrimidin 4amine Hydrochloride Abt 702, supplied by Tocris, 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/abt-702/n+cyclohexyl+2++3+5+dimethylpyrazol+1+yl++6+methylpyrimidin+4amine++cyppa+/pm39581530-83-1-43
    Average 90 stars, based on 1 article reviews
    5- (3-bromophenyl)-7-[6-(4-morpholinyl)-3-pyrido[2,3-d]byrimidin-4amine hydrochloride abt 702 - by Bioz Stars, 2026-09
    90/100 stars

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    Related Articles

    Injection:

    Article Title: An adenosinergic positive feedback loop extends pharmacological cardioprotection duration.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were orally administered ABT-702 by gavage of 3 mg kg 1 ABT-702 (Tocris Bioscience, Bristol, UK) or equivalent amounts of vehicle (DMSO) diluted in 100 μl saline.

    Article Title: Adenosine kinase (ADK) inhibition with ABT-702 induces ADK protein degradation and a distinct form of sustained cardioprotection.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were injected intraperitoneally (i.p.) with 10 mg/kg ABT-702 (Tocris Bioscience) or equivalent amounts of vehicle (DMSO) diluted in ~100 μl saline.

    Article Title: Adenosine kinase regulation of cardiomyocyte hypertrophy
    Article Snippet: Iodotubercidin (ITU) and ABT-702 were from Tocris (Ellisville, MO).

    Article Title: Mechanism of Activation of AMPK by Cordycepin
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Experimental Models: Cell Lines HepG2 Public Health England Cat# 85011430 RRID: CVCL_0027 U2OS TRex (Grumati et al., 2017) N/A Antibodies pT172 (AMPK-a) Cell Signaling Technology Cat# 2535; RRID: AB_331250 AMPK-a1 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a2 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a1 (for Western blotting) Cell Signaling Technology Cat# 2532; RRID: AB_330331 AMPK-a (pan-a, for Western blotting) AbCam Cat#ab32047; RRID: AB_722764 EZviewTM Red anti-FLAG M2 affinity gel Sigma-Aldrich Cat# F2426; RRID: AB_2616449 pACC1/pACC2 (S79/S212) Cell Signaling Technology Cat# 11818; RRID: AB_2687505 Streptavidin conjugated to 800 nm fluorophore (for detection of total ACC) Rockland Immunochemicals Cat# S000-32; pS792 (Raptor) Cell Signaling Technology Cat# 2083; RRID: AB_2249475 Raptor Cell Signaling Technology Cat# 2280; RRID: AB_561245 FLAG Sigma-Aldrich Cat# F2426; RRID: AB_2616449 Molecular Biology Kits KOD Hot Start kit Sigma-Aldrich Cat# 71842 MTT assay kit Abcam Cat# ab211091 PierceTM Protein Concentrator PES, 3K MWCO, 0.5 mL ThermoFisher Scientific Cat# 88512 Chemicals, Peptides, and Recombinant Proteins 2,4-dinitrophenol Sigma-Aldrich Cat# D198501 A-769662 (Goransson et al., 2007) N/A ABT-702 Tocris Cat# 2372 AICAR Abcam Cat# ab120358 AMPK purified from rat liver (Gowans et al., 2013) N/A antimycin A Sigma-Aldrich Cat# A8674 apyrase Sigma-Aldrich Cat# A6535 cordycepin Sigma-Aldrich Cat# C3394 cordycepin triphosphate Sigma-Aldrich Cat# C9137 dipyrimadole Tocris Cat# 0681 DMEM base Sigma-Aldrich Cat# D5030 Fugene 6 Promega Cat# E2691 Human AMPK (a1b2g1, bacterially expressed) (Hawley et al., 2012) N/A Human LKB1:STRAD-a:MO25-a complex (Zeqiraj et al., 2009) N/A Human PP2Ca (Davies et al., 1995) N/A phenformin Sigma-Aldrich Cat# P7045 rotenone Sigma-Aldrich Cat# R8875 Cell Chemical Biology 27, 214–222.e1–e4, February 20, 2020 e1

    Saline:

    Article Title: An adenosinergic positive feedback loop extends pharmacological cardioprotection duration.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were orally administered ABT-702 by gavage of 3 mg kg 1 ABT-702 (Tocris Bioscience, Bristol, UK) or equivalent amounts of vehicle (DMSO) diluted in 100 μl saline.

    Article Title: Adenosine kinase (ADK) inhibition with ABT-702 induces ADK protein degradation and a distinct form of sustained cardioprotection.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were injected intraperitoneally (i.p.) with 10 mg/kg ABT-702 (Tocris Bioscience) or equivalent amounts of vehicle (DMSO) diluted in ~100 μl saline.

    Article Title: Adenosine kinase regulation of cardiomyocyte hypertrophy
    Article Snippet: Iodotubercidin (ITU) and ABT-702 were from Tocris (Ellisville, MO).

    Article Title: Mechanism of Activation of AMPK by Cordycepin
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Experimental Models: Cell Lines HepG2 Public Health England Cat# 85011430 RRID: CVCL_0027 U2OS TRex (Grumati et al., 2017) N/A Antibodies pT172 (AMPK-a) Cell Signaling Technology Cat# 2535; RRID: AB_331250 AMPK-a1 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a2 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a1 (for Western blotting) Cell Signaling Technology Cat# 2532; RRID: AB_330331 AMPK-a (pan-a, for Western blotting) AbCam Cat#ab32047; RRID: AB_722764 EZviewTM Red anti-FLAG M2 affinity gel Sigma-Aldrich Cat# F2426; RRID: AB_2616449 pACC1/pACC2 (S79/S212) Cell Signaling Technology Cat# 11818; RRID: AB_2687505 Streptavidin conjugated to 800 nm fluorophore (for detection of total ACC) Rockland Immunochemicals Cat# S000-32; pS792 (Raptor) Cell Signaling Technology Cat# 2083; RRID: AB_2249475 Raptor Cell Signaling Technology Cat# 2280; RRID: AB_561245 FLAG Sigma-Aldrich Cat# F2426; RRID: AB_2616449 Molecular Biology Kits KOD Hot Start kit Sigma-Aldrich Cat# 71842 MTT assay kit Abcam Cat# ab211091 PierceTM Protein Concentrator PES, 3K MWCO, 0.5 mL ThermoFisher Scientific Cat# 88512 Chemicals, Peptides, and Recombinant Proteins 2,4-dinitrophenol Sigma-Aldrich Cat# D198501 A-769662 (Goransson et al., 2007) N/A ABT-702 Tocris Cat# 2372 AICAR Abcam Cat# ab120358 AMPK purified from rat liver (Gowans et al., 2013) N/A antimycin A Sigma-Aldrich Cat# A8674 apyrase Sigma-Aldrich Cat# A6535 cordycepin Sigma-Aldrich Cat# C3394 cordycepin triphosphate Sigma-Aldrich Cat# C9137 dipyrimadole Tocris Cat# 0681 DMEM base Sigma-Aldrich Cat# D5030 Fugene 6 Promega Cat# E2691 Human AMPK (a1b2g1, bacterially expressed) (Hawley et al., 2012) N/A Human LKB1:STRAD-a:MO25-a complex (Zeqiraj et al., 2009) N/A Human PP2Ca (Davies et al., 1995) N/A phenformin Sigma-Aldrich Cat# P7045 rotenone Sigma-Aldrich Cat# R8875 Cell Chemical Biology 27, 214–222.e1–e4, February 20, 2020 e1

    Immunoprecipitation:

    Article Title: An adenosinergic positive feedback loop extends pharmacological cardioprotection duration.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were orally administered ABT-702 by gavage of 3 mg kg 1 ABT-702 (Tocris Bioscience, Bristol, UK) or equivalent amounts of vehicle (DMSO) diluted in 100 μl saline.

    Article Title: Adenosine kinase (ADK) inhibition with ABT-702 induces ADK protein degradation and a distinct form of sustained cardioprotection.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were injected intraperitoneally (i.p.) with 10 mg/kg ABT-702 (Tocris Bioscience) or equivalent amounts of vehicle (DMSO) diluted in ~100 μl saline.

    Article Title: Adenosine kinase regulation of cardiomyocyte hypertrophy
    Article Snippet: Iodotubercidin (ITU) and ABT-702 were from Tocris (Ellisville, MO).

    Article Title: Mechanism of Activation of AMPK by Cordycepin
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Experimental Models: Cell Lines HepG2 Public Health England Cat# 85011430 RRID: CVCL_0027 U2OS TRex (Grumati et al., 2017) N/A Antibodies pT172 (AMPK-a) Cell Signaling Technology Cat# 2535; RRID: AB_331250 AMPK-a1 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a2 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a1 (for Western blotting) Cell Signaling Technology Cat# 2532; RRID: AB_330331 AMPK-a (pan-a, for Western blotting) AbCam Cat#ab32047; RRID: AB_722764 EZviewTM Red anti-FLAG M2 affinity gel Sigma-Aldrich Cat# F2426; RRID: AB_2616449 pACC1/pACC2 (S79/S212) Cell Signaling Technology Cat# 11818; RRID: AB_2687505 Streptavidin conjugated to 800 nm fluorophore (for detection of total ACC) Rockland Immunochemicals Cat# S000-32; pS792 (Raptor) Cell Signaling Technology Cat# 2083; RRID: AB_2249475 Raptor Cell Signaling Technology Cat# 2280; RRID: AB_561245 FLAG Sigma-Aldrich Cat# F2426; RRID: AB_2616449 Molecular Biology Kits KOD Hot Start kit Sigma-Aldrich Cat# 71842 MTT assay kit Abcam Cat# ab211091 PierceTM Protein Concentrator PES, 3K MWCO, 0.5 mL ThermoFisher Scientific Cat# 88512 Chemicals, Peptides, and Recombinant Proteins 2,4-dinitrophenol Sigma-Aldrich Cat# D198501 A-769662 (Goransson et al., 2007) N/A ABT-702 Tocris Cat# 2372 AICAR Abcam Cat# ab120358 AMPK purified from rat liver (Gowans et al., 2013) N/A antimycin A Sigma-Aldrich Cat# A8674 apyrase Sigma-Aldrich Cat# A6535 cordycepin Sigma-Aldrich Cat# C3394 cordycepin triphosphate Sigma-Aldrich Cat# C9137 dipyrimadole Tocris Cat# 0681 DMEM base Sigma-Aldrich Cat# D5030 Fugene 6 Promega Cat# E2691 Human AMPK (a1b2g1, bacterially expressed) (Hawley et al., 2012) N/A Human LKB1:STRAD-a:MO25-a complex (Zeqiraj et al., 2009) N/A Human PP2Ca (Davies et al., 1995) N/A phenformin Sigma-Aldrich Cat# P7045 rotenone Sigma-Aldrich Cat# R8875 Cell Chemical Biology 27, 214–222.e1–e4, February 20, 2020 e1

    Western Blot:

    Article Title: An adenosinergic positive feedback loop extends pharmacological cardioprotection duration.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were orally administered ABT-702 by gavage of 3 mg kg 1 ABT-702 (Tocris Bioscience, Bristol, UK) or equivalent amounts of vehicle (DMSO) diluted in 100 μl saline.

    Article Title: Adenosine kinase (ADK) inhibition with ABT-702 induces ADK protein degradation and a distinct form of sustained cardioprotection.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were injected intraperitoneally (i.p.) with 10 mg/kg ABT-702 (Tocris Bioscience) or equivalent amounts of vehicle (DMSO) diluted in ~100 μl saline.

    Article Title: Adenosine kinase regulation of cardiomyocyte hypertrophy
    Article Snippet: Iodotubercidin (ITU) and ABT-702 were from Tocris (Ellisville, MO).

    Article Title: Mechanism of Activation of AMPK by Cordycepin
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Experimental Models: Cell Lines HepG2 Public Health England Cat# 85011430 RRID: CVCL_0027 U2OS TRex (Grumati et al., 2017) N/A Antibodies pT172 (AMPK-a) Cell Signaling Technology Cat# 2535; RRID: AB_331250 AMPK-a1 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a2 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a1 (for Western blotting) Cell Signaling Technology Cat# 2532; RRID: AB_330331 AMPK-a (pan-a, for Western blotting) AbCam Cat#ab32047; RRID: AB_722764 EZviewTM Red anti-FLAG M2 affinity gel Sigma-Aldrich Cat# F2426; RRID: AB_2616449 pACC1/pACC2 (S79/S212) Cell Signaling Technology Cat# 11818; RRID: AB_2687505 Streptavidin conjugated to 800 nm fluorophore (for detection of total ACC) Rockland Immunochemicals Cat# S000-32; pS792 (Raptor) Cell Signaling Technology Cat# 2083; RRID: AB_2249475 Raptor Cell Signaling Technology Cat# 2280; RRID: AB_561245 FLAG Sigma-Aldrich Cat# F2426; RRID: AB_2616449 Molecular Biology Kits KOD Hot Start kit Sigma-Aldrich Cat# 71842 MTT assay kit Abcam Cat# ab211091 PierceTM Protein Concentrator PES, 3K MWCO, 0.5 mL ThermoFisher Scientific Cat# 88512 Chemicals, Peptides, and Recombinant Proteins 2,4-dinitrophenol Sigma-Aldrich Cat# D198501 A-769662 (Goransson et al., 2007) N/A ABT-702 Tocris Cat# 2372 AICAR Abcam Cat# ab120358 AMPK purified from rat liver (Gowans et al., 2013) N/A antimycin A Sigma-Aldrich Cat# A8674 apyrase Sigma-Aldrich Cat# A6535 cordycepin Sigma-Aldrich Cat# C3394 cordycepin triphosphate Sigma-Aldrich Cat# C9137 dipyrimadole Tocris Cat# 0681 DMEM base Sigma-Aldrich Cat# D5030 Fugene 6 Promega Cat# E2691 Human AMPK (a1b2g1, bacterially expressed) (Hawley et al., 2012) N/A Human LKB1:STRAD-a:MO25-a complex (Zeqiraj et al., 2009) N/A Human PP2Ca (Davies et al., 1995) N/A phenformin Sigma-Aldrich Cat# P7045 rotenone Sigma-Aldrich Cat# R8875 Cell Chemical Biology 27, 214–222.e1–e4, February 20, 2020 e1

    MTT Assay:

    Article Title: An adenosinergic positive feedback loop extends pharmacological cardioprotection duration.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were orally administered ABT-702 by gavage of 3 mg kg 1 ABT-702 (Tocris Bioscience, Bristol, UK) or equivalent amounts of vehicle (DMSO) diluted in 100 μl saline.

    Article Title: Adenosine kinase (ADK) inhibition with ABT-702 induces ADK protein degradation and a distinct form of sustained cardioprotection.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were injected intraperitoneally (i.p.) with 10 mg/kg ABT-702 (Tocris Bioscience) or equivalent amounts of vehicle (DMSO) diluted in ~100 μl saline.

    Article Title: Adenosine kinase regulation of cardiomyocyte hypertrophy
    Article Snippet: Iodotubercidin (ITU) and ABT-702 were from Tocris (Ellisville, MO).

    Article Title: Mechanism of Activation of AMPK by Cordycepin
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Experimental Models: Cell Lines HepG2 Public Health England Cat# 85011430 RRID: CVCL_0027 U2OS TRex (Grumati et al., 2017) N/A Antibodies pT172 (AMPK-a) Cell Signaling Technology Cat# 2535; RRID: AB_331250 AMPK-a1 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a2 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a1 (for Western blotting) Cell Signaling Technology Cat# 2532; RRID: AB_330331 AMPK-a (pan-a, for Western blotting) AbCam Cat#ab32047; RRID: AB_722764 EZviewTM Red anti-FLAG M2 affinity gel Sigma-Aldrich Cat# F2426; RRID: AB_2616449 pACC1/pACC2 (S79/S212) Cell Signaling Technology Cat# 11818; RRID: AB_2687505 Streptavidin conjugated to 800 nm fluorophore (for detection of total ACC) Rockland Immunochemicals Cat# S000-32; pS792 (Raptor) Cell Signaling Technology Cat# 2083; RRID: AB_2249475 Raptor Cell Signaling Technology Cat# 2280; RRID: AB_561245 FLAG Sigma-Aldrich Cat# F2426; RRID: AB_2616449 Molecular Biology Kits KOD Hot Start kit Sigma-Aldrich Cat# 71842 MTT assay kit Abcam Cat# ab211091 PierceTM Protein Concentrator PES, 3K MWCO, 0.5 mL ThermoFisher Scientific Cat# 88512 Chemicals, Peptides, and Recombinant Proteins 2,4-dinitrophenol Sigma-Aldrich Cat# D198501 A-769662 (Goransson et al., 2007) N/A ABT-702 Tocris Cat# 2372 AICAR Abcam Cat# ab120358 AMPK purified from rat liver (Gowans et al., 2013) N/A antimycin A Sigma-Aldrich Cat# A8674 apyrase Sigma-Aldrich Cat# A6535 cordycepin Sigma-Aldrich Cat# C3394 cordycepin triphosphate Sigma-Aldrich Cat# C9137 dipyrimadole Tocris Cat# 0681 DMEM base Sigma-Aldrich Cat# D5030 Fugene 6 Promega Cat# E2691 Human AMPK (a1b2g1, bacterially expressed) (Hawley et al., 2012) N/A Human LKB1:STRAD-a:MO25-a complex (Zeqiraj et al., 2009) N/A Human PP2Ca (Davies et al., 1995) N/A phenformin Sigma-Aldrich Cat# P7045 rotenone Sigma-Aldrich Cat# R8875 Cell Chemical Biology 27, 214–222.e1–e4, February 20, 2020 e1

    Recombinant:

    Article Title: An adenosinergic positive feedback loop extends pharmacological cardioprotection duration.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were orally administered ABT-702 by gavage of 3 mg kg 1 ABT-702 (Tocris Bioscience, Bristol, UK) or equivalent amounts of vehicle (DMSO) diluted in 100 μl saline.

    Article Title: Adenosine kinase (ADK) inhibition with ABT-702 induces ADK protein degradation and a distinct form of sustained cardioprotection.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were injected intraperitoneally (i.p.) with 10 mg/kg ABT-702 (Tocris Bioscience) or equivalent amounts of vehicle (DMSO) diluted in ~100 μl saline.

    Article Title: Adenosine kinase regulation of cardiomyocyte hypertrophy
    Article Snippet: Iodotubercidin (ITU) and ABT-702 were from Tocris (Ellisville, MO).

    Article Title: Mechanism of Activation of AMPK by Cordycepin
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Experimental Models: Cell Lines HepG2 Public Health England Cat# 85011430 RRID: CVCL_0027 U2OS TRex (Grumati et al., 2017) N/A Antibodies pT172 (AMPK-a) Cell Signaling Technology Cat# 2535; RRID: AB_331250 AMPK-a1 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a2 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a1 (for Western blotting) Cell Signaling Technology Cat# 2532; RRID: AB_330331 AMPK-a (pan-a, for Western blotting) AbCam Cat#ab32047; RRID: AB_722764 EZviewTM Red anti-FLAG M2 affinity gel Sigma-Aldrich Cat# F2426; RRID: AB_2616449 pACC1/pACC2 (S79/S212) Cell Signaling Technology Cat# 11818; RRID: AB_2687505 Streptavidin conjugated to 800 nm fluorophore (for detection of total ACC) Rockland Immunochemicals Cat# S000-32; pS792 (Raptor) Cell Signaling Technology Cat# 2083; RRID: AB_2249475 Raptor Cell Signaling Technology Cat# 2280; RRID: AB_561245 FLAG Sigma-Aldrich Cat# F2426; RRID: AB_2616449 Molecular Biology Kits KOD Hot Start kit Sigma-Aldrich Cat# 71842 MTT assay kit Abcam Cat# ab211091 PierceTM Protein Concentrator PES, 3K MWCO, 0.5 mL ThermoFisher Scientific Cat# 88512 Chemicals, Peptides, and Recombinant Proteins 2,4-dinitrophenol Sigma-Aldrich Cat# D198501 A-769662 (Goransson et al., 2007) N/A ABT-702 Tocris Cat# 2372 AICAR Abcam Cat# ab120358 AMPK purified from rat liver (Gowans et al., 2013) N/A antimycin A Sigma-Aldrich Cat# A8674 apyrase Sigma-Aldrich Cat# A6535 cordycepin Sigma-Aldrich Cat# C3394 cordycepin triphosphate Sigma-Aldrich Cat# C9137 dipyrimadole Tocris Cat# 0681 DMEM base Sigma-Aldrich Cat# D5030 Fugene 6 Promega Cat# E2691 Human AMPK (a1b2g1, bacterially expressed) (Hawley et al., 2012) N/A Human LKB1:STRAD-a:MO25-a complex (Zeqiraj et al., 2009) N/A Human PP2Ca (Davies et al., 1995) N/A phenformin Sigma-Aldrich Cat# P7045 rotenone Sigma-Aldrich Cat# R8875 Cell Chemical Biology 27, 214–222.e1–e4, February 20, 2020 e1

    Purification:

    Article Title: An adenosinergic positive feedback loop extends pharmacological cardioprotection duration.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were orally administered ABT-702 by gavage of 3 mg kg 1 ABT-702 (Tocris Bioscience, Bristol, UK) or equivalent amounts of vehicle (DMSO) diluted in 100 μl saline.

    Article Title: Adenosine kinase (ADK) inhibition with ABT-702 induces ADK protein degradation and a distinct form of sustained cardioprotection.
    Article Snippet: At 16–20 weeks of age, randomly assigned mice were injected intraperitoneally (i.p.) with 10 mg/kg ABT-702 (Tocris Bioscience) or equivalent amounts of vehicle (DMSO) diluted in ~100 μl saline.

    Article Title: Adenosine kinase regulation of cardiomyocyte hypertrophy
    Article Snippet: Iodotubercidin (ITU) and ABT-702 were from Tocris (Ellisville, MO).

    Article Title: Mechanism of Activation of AMPK by Cordycepin
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Experimental Models: Cell Lines HepG2 Public Health England Cat# 85011430 RRID: CVCL_0027 U2OS TRex (Grumati et al., 2017) N/A Antibodies pT172 (AMPK-a) Cell Signaling Technology Cat# 2535; RRID: AB_331250 AMPK-a1 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a2 (for immunoprecipitation) (Woods et al., 1996) N/A AMPK-a1 (for Western blotting) Cell Signaling Technology Cat# 2532; RRID: AB_330331 AMPK-a (pan-a, for Western blotting) AbCam Cat#ab32047; RRID: AB_722764 EZviewTM Red anti-FLAG M2 affinity gel Sigma-Aldrich Cat# F2426; RRID: AB_2616449 pACC1/pACC2 (S79/S212) Cell Signaling Technology Cat# 11818; RRID: AB_2687505 Streptavidin conjugated to 800 nm fluorophore (for detection of total ACC) Rockland Immunochemicals Cat# S000-32; pS792 (Raptor) Cell Signaling Technology Cat# 2083; RRID: AB_2249475 Raptor Cell Signaling Technology Cat# 2280; RRID: AB_561245 FLAG Sigma-Aldrich Cat# F2426; RRID: AB_2616449 Molecular Biology Kits KOD Hot Start kit Sigma-Aldrich Cat# 71842 MTT assay kit Abcam Cat# ab211091 PierceTM Protein Concentrator PES, 3K MWCO, 0.5 mL ThermoFisher Scientific Cat# 88512 Chemicals, Peptides, and Recombinant Proteins 2,4-dinitrophenol Sigma-Aldrich Cat# D198501 A-769662 (Goransson et al., 2007) N/A ABT-702 Tocris Cat# 2372 AICAR Abcam Cat# ab120358 AMPK purified from rat liver (Gowans et al., 2013) N/A antimycin A Sigma-Aldrich Cat# A8674 apyrase Sigma-Aldrich Cat# A6535 cordycepin Sigma-Aldrich Cat# C3394 cordycepin triphosphate Sigma-Aldrich Cat# C9137 dipyrimadole Tocris Cat# 0681 DMEM base Sigma-Aldrich Cat# D5030 Fugene 6 Promega Cat# E2691 Human AMPK (a1b2g1, bacterially expressed) (Hawley et al., 2012) N/A Human LKB1:STRAD-a:MO25-a complex (Zeqiraj et al., 2009) N/A Human PP2Ca (Davies et al., 1995) N/A phenformin Sigma-Aldrich Cat# P7045 rotenone Sigma-Aldrich Cat# R8875 Cell Chemical Biology 27, 214–222.e1–e4, February 20, 2020 e1



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    Effects of inhibitors of adenosine metabolism on the utilization of adenosine for ATP restoration in glucose-fed astrocytes. Astrocyte cultures had been preincubated for 60 min in glucose-free IB with 1 µM of BAM15 to lower the cellular ATP content before the cells were incubated in glucose (5 mM)-containing IB without or with 30 µM adenosine in the absence or the presence of the adenosine kinase inhibitor ABT-702 (ABT; 10 µM), the adenosine deaminase inhibitor DCF (1 µM) and/or the purine nucleoside phosphorylase inhibitor forodesine (Foro; 10 µM). After 60 min, the cellular ATP content ( a ) and the extracellular LDH activity ( b ) were determined. The data shown are means ± SD of values obtained in three experiments performed on independently prepared cultures. The average initial ATP content of the cultures (36.8 ± 1.6 nmol/mg) is indicated by the black dotted line in panel a. The protein content of the cultures was 130 ± 12 µg/well and the initial cellular LDH activity 119 ± 5 nmol/(min × well). The significance of differences (ANOVA) compared with the data obtained for the respective control incubation (absence of all inhibitors) is indicated by *** p < 0.001. The significance of differences (t-test) between data for adenosine-free and the respective adenosine-containing incubations is indicated by # p < 0.05, ## p < 0.01and ### p < 0.001

    Journal: Neurochemical Research

    Article Title: ATP Restoration by ATP-Deprived Cultured Primary Astrocytes

    doi: 10.1007/s11064-024-04276-9

    Figure Lengend Snippet: Effects of inhibitors of adenosine metabolism on the utilization of adenosine for ATP restoration in glucose-fed astrocytes. Astrocyte cultures had been preincubated for 60 min in glucose-free IB with 1 µM of BAM15 to lower the cellular ATP content before the cells were incubated in glucose (5 mM)-containing IB without or with 30 µM adenosine in the absence or the presence of the adenosine kinase inhibitor ABT-702 (ABT; 10 µM), the adenosine deaminase inhibitor DCF (1 µM) and/or the purine nucleoside phosphorylase inhibitor forodesine (Foro; 10 µM). After 60 min, the cellular ATP content ( a ) and the extracellular LDH activity ( b ) were determined. The data shown are means ± SD of values obtained in three experiments performed on independently prepared cultures. The average initial ATP content of the cultures (36.8 ± 1.6 nmol/mg) is indicated by the black dotted line in panel a. The protein content of the cultures was 130 ± 12 µg/well and the initial cellular LDH activity 119 ± 5 nmol/(min × well). The significance of differences (ANOVA) compared with the data obtained for the respective control incubation (absence of all inhibitors) is indicated by *** p < 0.001. The significance of differences (t-test) between data for adenosine-free and the respective adenosine-containing incubations is indicated by # p < 0.05, ## p < 0.01and ### p < 0.001

    Article Snippet: The adenosine kinase inhibitor ABT-702 and the purine nucleoside phosphorylase inhibitor forodesine hydrochloride were obtained from MedChemExpress (Monmouth Junction, NJ, USA).

    Techniques: Incubation, Activity Assay, Control

    Prevention of the exclusive utilization of adenosine for ATP restoration by inhibitors of adenosine metabolism. Astrocyte cultures had been preincubated for 60 min in glucose-free IB with 1 µM of BAM15 to lower the cellular ATP content before the cells were incubated in glucose-free IB with 100 µM adenosine in the absence or the presence of the adenosine kinase inhibitor ABT-702 (ABT; 10 µM), the adenosine deaminase inhibitor DCF (1 µM) and/or the purine nucleoside phosphorylase inhibitor forodesine (Foro; 10 µM). After 60 min, the cellular ATP content ( a ) and the extracellular LDH activity ( b ) were determined. The data shown are means ± SD of values obtained in three experiments performed on independently prepared cultures. The average initial ATP content of the cultures (37.3 ± 2.4 nmol/mg) is indicated by the black dotted line in panel a. The protein content of the cultures was 136 ± 22 µg/well and the initial cellular LDH activity was 163 ± 59 nmol/(min × well). The significance of differences (ANOVA) compared with the data obtained for the control incubation (absence of all inhibitors) is indicated by *** p < 0.001. The significance of differences (ANOVA) compared with the data obtained after the 60 min pre-incubation (8.1 ± 4.4 nmol/mg; indicated by the black dashed lines in panel a) is indicated by + p < 0.05 and +++ p < 0.001

    Journal: Neurochemical Research

    Article Title: ATP Restoration by ATP-Deprived Cultured Primary Astrocytes

    doi: 10.1007/s11064-024-04276-9

    Figure Lengend Snippet: Prevention of the exclusive utilization of adenosine for ATP restoration by inhibitors of adenosine metabolism. Astrocyte cultures had been preincubated for 60 min in glucose-free IB with 1 µM of BAM15 to lower the cellular ATP content before the cells were incubated in glucose-free IB with 100 µM adenosine in the absence or the presence of the adenosine kinase inhibitor ABT-702 (ABT; 10 µM), the adenosine deaminase inhibitor DCF (1 µM) and/or the purine nucleoside phosphorylase inhibitor forodesine (Foro; 10 µM). After 60 min, the cellular ATP content ( a ) and the extracellular LDH activity ( b ) were determined. The data shown are means ± SD of values obtained in three experiments performed on independently prepared cultures. The average initial ATP content of the cultures (37.3 ± 2.4 nmol/mg) is indicated by the black dotted line in panel a. The protein content of the cultures was 136 ± 22 µg/well and the initial cellular LDH activity was 163 ± 59 nmol/(min × well). The significance of differences (ANOVA) compared with the data obtained for the control incubation (absence of all inhibitors) is indicated by *** p < 0.001. The significance of differences (ANOVA) compared with the data obtained after the 60 min pre-incubation (8.1 ± 4.4 nmol/mg; indicated by the black dashed lines in panel a) is indicated by + p < 0.05 and +++ p < 0.001

    Article Snippet: The adenosine kinase inhibitor ABT-702 and the purine nucleoside phosphorylase inhibitor forodesine hydrochloride were obtained from MedChemExpress (Monmouth Junction, NJ, USA).

    Techniques: Incubation, Activity Assay, Control