e2 Search Results


prostaglandin e2 pge2 assay kit  (R&D Systems)
96
R&D Systems prostaglandin e2 pge2 assay kit
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
Prostaglandin E2 Pge2 Assay Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/e2/Prostaglandin+E2+Parameter+Assay+Kit/pm19106818-46-0-7
Average 96 stars, based on 1 article reviews
prostaglandin e2 pge2 assay kit - by Bioz Stars, 2026-10
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e2 elisa kit  (Elabscience Biotechnology)
95
Elabscience Biotechnology e2 elisa kit
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
E2 Elisa Kit, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/e2/QuicKey+Pro+Rat+E2+(Estradiol)+ELISA+Kit/pm40429851-419-8-12
Average 95 stars, based on 1 article reviews
e2 elisa kit - by Bioz Stars, 2026-10
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e 2 elisa kit  (Elabscience Biotechnology)
93
Elabscience Biotechnology e 2 elisa kit
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
E 2 Elisa Kit, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/e2/E2+(Estradiol)+ELISA+Kit/pmc11350841-66-28-43
Average 93 stars, based on 1 article reviews
e 2 elisa kit - by Bioz Stars, 2026-10
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prostaglandin e 2  (Tocris)
97
Tocris prostaglandin e 2
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
Prostaglandin E 2, supplied by Tocris, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/e2/Prostaglandin+E2/pmc10684765-112-53-59
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prostaglandin e 2 - by Bioz Stars, 2026-10
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prostaglandin e2  (Tocris)
96
Tocris prostaglandin e2
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
Prostaglandin E2, supplied by Tocris, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/e2/Prostaglandin+E2/10__1074_slash_jbc__m609695200-57-1-31
Average 96 stars, based on 1 article reviews
prostaglandin e2 - by Bioz Stars, 2026-10
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rat pge2  (Elabscience Biotechnology)
96
Elabscience Biotechnology rat pge2
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
Rat Pge2, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/e2/PGE2+(Prostaglandin+E2)+ELISA+Kit/10__1155_slash_jfbc_slash_6639397-75-19-37
Average 96 stars, based on 1 article reviews
rat pge2 - by Bioz Stars, 2026-10
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human recombinant serpine2  (R&D Systems)
93
R&D Systems human recombinant serpine2
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
Human Recombinant Serpine2, supplied by R&D Systems, 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/e2/Recombinant+Human+Serpin+E2%2FPN1+Protein%2C+CF/pm26305372-50-0-6
Average 93 stars, based on 1 article reviews
human recombinant serpine2 - by Bioz Stars, 2026-10
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e2 conjugase  (R&D Systems)
95
R&D Systems e2 conjugase
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
E2 Conjugase, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/e2/Recombinant+Human%2FMouse%2FRat+UbcH5c%2FUBE2D3+Protein%2C+CF/bio_rxiv__2021__03__17__435897-108-53-57
Average 95 stars, based on 1 article reviews
e2 conjugase - by Bioz Stars, 2026-10
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parametertm pge 2 immunoassay kit  (R&D Systems)
97
R&D Systems parametertm pge 2 immunoassay kit
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
Parametertm Pge 2 Immunoassay Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/e2/Prostaglandin+E2+Parameter+Assay+Kit/pmc03558384-108-28-33
Average 97 stars, based on 1 article reviews
parametertm pge 2 immunoassay kit - by Bioz Stars, 2026-10
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ubch7 ube2l3  (R&D Systems)
93
R&D Systems ubch7 ube2l3
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
Ubch7 Ube2l3, supplied by R&D Systems, 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/e2/Recombinant+Human%2FMouse%2FRat+UbcH7%2FUBE2L3+Protein%2C+CF/pmc05207158-1043-0-9
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human ubch2 ube2h myc ddk  (OriGene)
90
OriGene human ubch2 ube2h myc ddk
FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by <t>PGE2</t> ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.
Human Ubch2 Ube2h Myc Ddk, supplied by OriGene, 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/e2/UBE2H+(NM_003344)+Human+Tagged+ORF+Clone/pm33298911-421-1-7
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mouse ube2o  (OriGene)
90
OriGene mouse ube2o
(A) An RNA interference (RNAi) screen identifies <t>UBE2O</t> as a potent regulator of glucose uptake in myotubes. Primary normal human skeletal myotubes (HSkMs) and C2C12 mouse myotubes expressing a nontargeting control siRNA pool (Cont. siRNAs) or a synthetic siRNA library targeting 30 human and mouse E2s, respectively, were subjected to a screen for insulin-stimulated 2-deoxyglucose uptake. 2-DG6P, 2-deoxyglucose-6-phosphate. n = 3. (B) Total RNAs from quadriceps skeletal muscles of mice fed a normal chow or an HFD for 28 weeks were subjected to RT-qPCR. n = 4. (C and D) Lysates from quadriceps skeletal muscles of 9-month-old Ube2o+/+ mice fed normal chow or an HFD for 28 weeks (C) and 25-week-old type 2 diabetic db/db and lean (db/+) mice (D) were subjected to immunoblotting. Error bars represent ±SEM. P value was determined by Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001, control siRNAs HSkMs vs. E2 siRNAs HSkMs; #P < 0.05, ##P < 0.01, ###P < 0.001, control siRNAs C2C12 vs. E2 siRNAs C2C12; †P < 0.05, †††P < 0.001, chow vs. HFD. Uncropped gels and blots are available in the supplemental material.
Mouse Ube2o, supplied by OriGene, 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/e2/Ube2o+(BC059193)+Mouse+Tagged+ORF+Clone/pmc06629239-499-6-8
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Image Search Results


FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by PGE2 ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.

Journal: Shock (Augusta, Ga.)

Article Title: Evodiamine represses hypoxia-induced inflammatory proteins expression and hypoxia-inducible factor 1alpha accumulation in RAW264.7.

doi: 10.1097/SHK.0b013e31819940cb

Figure Lengend Snippet: FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by PGE2 ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.

Article Snippet: Prostaglandin E2 (PGE2) assay kit was from R&D Systems (Minneapolis, Minn).

Techniques: Incubation, Concentration Assay, Nucleic Acid Electrophoresis, Western Blot, Enzyme-linked Immunosorbent Assay, Sulforhodamine B Assay

(A) An RNA interference (RNAi) screen identifies UBE2O as a potent regulator of glucose uptake in myotubes. Primary normal human skeletal myotubes (HSkMs) and C2C12 mouse myotubes expressing a nontargeting control siRNA pool (Cont. siRNAs) or a synthetic siRNA library targeting 30 human and mouse E2s, respectively, were subjected to a screen for insulin-stimulated 2-deoxyglucose uptake. 2-DG6P, 2-deoxyglucose-6-phosphate. n = 3. (B) Total RNAs from quadriceps skeletal muscles of mice fed a normal chow or an HFD for 28 weeks were subjected to RT-qPCR. n = 4. (C and D) Lysates from quadriceps skeletal muscles of 9-month-old Ube2o+/+ mice fed normal chow or an HFD for 28 weeks (C) and 25-week-old type 2 diabetic db/db and lean (db/+) mice (D) were subjected to immunoblotting. Error bars represent ±SEM. P value was determined by Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001, control siRNAs HSkMs vs. E2 siRNAs HSkMs; #P < 0.05, ##P < 0.01, ###P < 0.001, control siRNAs C2C12 vs. E2 siRNAs C2C12; †P < 0.05, †††P < 0.001, chow vs. HFD. Uncropped gels and blots are available in the supplemental material.

Journal: JCI Insight

Article Title: A muscle-specific UBE2O/AMPK α 2 axis promotes insulin resistance and metabolic syndrome in obesity

doi: 10.1172/jci.insight.128269

Figure Lengend Snippet: (A) An RNA interference (RNAi) screen identifies UBE2O as a potent regulator of glucose uptake in myotubes. Primary normal human skeletal myotubes (HSkMs) and C2C12 mouse myotubes expressing a nontargeting control siRNA pool (Cont. siRNAs) or a synthetic siRNA library targeting 30 human and mouse E2s, respectively, were subjected to a screen for insulin-stimulated 2-deoxyglucose uptake. 2-DG6P, 2-deoxyglucose-6-phosphate. n = 3. (B) Total RNAs from quadriceps skeletal muscles of mice fed a normal chow or an HFD for 28 weeks were subjected to RT-qPCR. n = 4. (C and D) Lysates from quadriceps skeletal muscles of 9-month-old Ube2o+/+ mice fed normal chow or an HFD for 28 weeks (C) and 25-week-old type 2 diabetic db/db and lean (db/+) mice (D) were subjected to immunoblotting. Error bars represent ±SEM. P value was determined by Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001, control siRNAs HSkMs vs. E2 siRNAs HSkMs; #P < 0.05, ##P < 0.01, ###P < 0.001, control siRNAs C2C12 vs. E2 siRNAs C2C12; †P < 0.05, †††P < 0.001, chow vs. HFD. Uncropped gels and blots are available in the supplemental material.

Article Snippet: The mutant construct (C1037S) of Flag-tagged mouse UBE2O (Origene) was generated by site-directed mutagenesis and cloned into pBabe retroviral vector.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

(A) Body weight increase of Ube2o+/+ and Ube2o–/– mice fed an HFD for 21 weeks. Ube2o+/+ n = 7, Ube2o–/– n = 9. (B) Food intake (grams per day) in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Ube2o+/+ n = 6, Ube2o–/– n = 8. (C) Fat mass of Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Ube2o+/+ n = 5, Ube2o–/– n = 9. (D) Adipose depot weight of Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. SAT, subcutaneous adipose tissue; VAT, visceral adipose tissue; BAT, brown adipose tissue. Ube2o+/+ n = 6, Ube2o–/– n = 9. (E) H&E-stained sections of SAT in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Scale bars: 75 μm. (F) Plasma levels of total cholesterol (TC) (mg/dL), free fatty acid (FFA; mEq/L), HDL (mg/dL), LDL (mg/dL) in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. n = 6. (G) Liver weight of Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Ube2o+/+ n = 6, Ube2o–/– n = 9. (H) Hepatic triglyceride (TG) level per gram of liver in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. n = 6. (I) H&E-stained sections of liver in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Scale bars: 75 μm. (J) Oxygen consumption (VO2) per kg lean mass of Ube2o+/+ and Ube2o–/– mice on an HFD for 20 weeks was determined in metabolic chambers. Ube2o+/+ n = 6, Ube2o–/– n = 9. (K) Respiratory exchange ratio (RER; VCO2/VO2) of Ube2o+/+ and Ube2o–/– mice on an HFD for 20 weeks was determined in metabolic chambers. Ube2o+/+ n = 6, Ube2o–/– n = 9. (L) Energy expenditure during day and night per kg of lean mass of Ube2o+/+ and Ube2o–/– mice on an HFD for 20 weeks was determined in metabolic chambers. Ube2o+/+ n = 6, Ube2o–/– n = 9. (M) Home cage locomotor activity during both the light and dark phases of Ube2o+/+ and Ube2o–/– mice on an HFD for 20 weeks was determined by using an automated combined indirect calorimetry system. Ube2o+/+ n = 5, Ube2o–/– n = 8. (N) Rectal temperature of Ube2o+/+ and Ube2o–/– mice on an HFD for 12 weeks. Ube2o+/+ n = 8, Ube2o–/– n = 9. Error bars represent ±SEM. P value was determined by Student’s t test (*P < 0.05, **P < 0.01, ***P < 0.001).

Journal: JCI Insight

Article Title: A muscle-specific UBE2O/AMPK α 2 axis promotes insulin resistance and metabolic syndrome in obesity

doi: 10.1172/jci.insight.128269

Figure Lengend Snippet: (A) Body weight increase of Ube2o+/+ and Ube2o–/– mice fed an HFD for 21 weeks. Ube2o+/+ n = 7, Ube2o–/– n = 9. (B) Food intake (grams per day) in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Ube2o+/+ n = 6, Ube2o–/– n = 8. (C) Fat mass of Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Ube2o+/+ n = 5, Ube2o–/– n = 9. (D) Adipose depot weight of Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. SAT, subcutaneous adipose tissue; VAT, visceral adipose tissue; BAT, brown adipose tissue. Ube2o+/+ n = 6, Ube2o–/– n = 9. (E) H&E-stained sections of SAT in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Scale bars: 75 μm. (F) Plasma levels of total cholesterol (TC) (mg/dL), free fatty acid (FFA; mEq/L), HDL (mg/dL), LDL (mg/dL) in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. n = 6. (G) Liver weight of Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Ube2o+/+ n = 6, Ube2o–/– n = 9. (H) Hepatic triglyceride (TG) level per gram of liver in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. n = 6. (I) H&E-stained sections of liver in Ube2o+/+ and Ube2o–/– mice on an HFD for 28 weeks. Scale bars: 75 μm. (J) Oxygen consumption (VO2) per kg lean mass of Ube2o+/+ and Ube2o–/– mice on an HFD for 20 weeks was determined in metabolic chambers. Ube2o+/+ n = 6, Ube2o–/– n = 9. (K) Respiratory exchange ratio (RER; VCO2/VO2) of Ube2o+/+ and Ube2o–/– mice on an HFD for 20 weeks was determined in metabolic chambers. Ube2o+/+ n = 6, Ube2o–/– n = 9. (L) Energy expenditure during day and night per kg of lean mass of Ube2o+/+ and Ube2o–/– mice on an HFD for 20 weeks was determined in metabolic chambers. Ube2o+/+ n = 6, Ube2o–/– n = 9. (M) Home cage locomotor activity during both the light and dark phases of Ube2o+/+ and Ube2o–/– mice on an HFD for 20 weeks was determined by using an automated combined indirect calorimetry system. Ube2o+/+ n = 5, Ube2o–/– n = 8. (N) Rectal temperature of Ube2o+/+ and Ube2o–/– mice on an HFD for 12 weeks. Ube2o+/+ n = 8, Ube2o–/– n = 9. Error bars represent ±SEM. P value was determined by Student’s t test (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: The mutant construct (C1037S) of Flag-tagged mouse UBE2O (Origene) was generated by site-directed mutagenesis and cloned into pBabe retroviral vector.

Techniques: Staining, Activity Assay

(A) Blood glucose was measured Ube2o+/+ and Ube2o–/– mice on an HFD for 21 weeks. n = 6. (B and C) Glucose (GTT) (B) and insulin (ITT) (C) tolerance tests in Ube2o+/+ and Ube2o–/– mice on HFD for 21~22 weeks. Insets indicate AUC (B) and area above the curve (AAC) (C). Insulinemia at 15 minutes after the injection of glucose during a GTT test is shown in the right panel (B). Ube2o+/+ n = 6, Ube2o–/– n = 9. (D–F) Rates of basal and clamp endogenous glucose production (D), glucose infusion (E), and glucose disposal rate (F) during a hyperinsulinemic-euglycemic clamp study in Ube2o+/+ and Ube2o–/– mice. n = 6. (G) Insulin-stimulated 14C-2-deoxyglucose uptake was assessed in visceral WAT and gastrocnemius muscle of Ube2o+/+ and Ube2o–/– mice during the final 45 minutes of the hyperinsulinemic-euglycemic clamps. n = 7. (H) Immunoblots (left) and statistical data (right) showing insulin-induced (200 nM) tyrosine phosphorylation of IRS1 immunoprecipitates and S473 phosphorylation of AKT protein and their total protein levels in the extensor digitorum longus muscle from Ube2o+/+ and Ube2o–/– mice on an HFD. n = 3. Error bars represent ± =SEM. P value was determined by Student’s t test (*P < 0.05, **P < 0.01, ***P < 0.001).

Journal: JCI Insight

Article Title: A muscle-specific UBE2O/AMPK α 2 axis promotes insulin resistance and metabolic syndrome in obesity

doi: 10.1172/jci.insight.128269

Figure Lengend Snippet: (A) Blood glucose was measured Ube2o+/+ and Ube2o–/– mice on an HFD for 21 weeks. n = 6. (B and C) Glucose (GTT) (B) and insulin (ITT) (C) tolerance tests in Ube2o+/+ and Ube2o–/– mice on HFD for 21~22 weeks. Insets indicate AUC (B) and area above the curve (AAC) (C). Insulinemia at 15 minutes after the injection of glucose during a GTT test is shown in the right panel (B). Ube2o+/+ n = 6, Ube2o–/– n = 9. (D–F) Rates of basal and clamp endogenous glucose production (D), glucose infusion (E), and glucose disposal rate (F) during a hyperinsulinemic-euglycemic clamp study in Ube2o+/+ and Ube2o–/– mice. n = 6. (G) Insulin-stimulated 14C-2-deoxyglucose uptake was assessed in visceral WAT and gastrocnemius muscle of Ube2o+/+ and Ube2o–/– mice during the final 45 minutes of the hyperinsulinemic-euglycemic clamps. n = 7. (H) Immunoblots (left) and statistical data (right) showing insulin-induced (200 nM) tyrosine phosphorylation of IRS1 immunoprecipitates and S473 phosphorylation of AKT protein and their total protein levels in the extensor digitorum longus muscle from Ube2o+/+ and Ube2o–/– mice on an HFD. n = 3. Error bars represent ± =SEM. P value was determined by Student’s t test (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: The mutant construct (C1037S) of Flag-tagged mouse UBE2O (Origene) was generated by site-directed mutagenesis and cloned into pBabe retroviral vector.

Techniques: Injection, Western Blot

(A) UBE2O (top) or AMPKα2 (bottom) immunoprecipitates of lysates from skeletal muscle of control (Ube2ofl/fl) and Ube2oΔMus mice were subjected to immunoblotting for AMPKα2 or UBE2O. (B) Lysates from C2C12 myotubes transfected with the indicated plasmids were subjected to metal affinity purification for His-tagged ubiquitin (His-Ub), then immunoblotting for ubiquitinated AMPKα2. Ni-NTA, Ni2+-nitrilotriacetic acid. (C) Recombinant AMPKα2 proteins were subjected to in vitro ubiquitination assay in the presence of in vitro translated WT or C1037S (CS) mutant UBE2O. (D) Lysates from C2C12 myotubes expressing WT or CS mutant UBE2O treated with MG132 (10 μM) for 6 hours were subjected to immunoblotting. (E) Lysates from C2C12 myotubes expressing WT or CS mutant UBE2O treated with cycloheximide (CHX) for the indicated times were subjected to immunoblotting (top). AMPKα2 or AMPKα1 protein levels were quantified by normalizing to the intensity of the HSP90 band (bottom). (F) Lysates from skeletal muscle of 12-week-old Ube2o+/+, Ube2o–/–, Ube2ofl/fl, and Ube2oΔMus mice were subjected to immunoblotting for the indicated proteins. (G) Lysates from C2C12 myotubes expressing UBE2O shRNA together with exogenous WT or CS mutant UBE2O from a Ube2o ORF transcript lacking the 3′ UTR sequence targeted by shRNA (shUbe2o-3′UTR) were subjected to immunoblotting for the indicated proteins. (H) Insulin-stimulated glucose uptake rate of C2C12 myotubes from G. 2-DG6P, 2-deoxyglucose-6-phosphate. n = 4. (I) Lysates from C2C12 myotubes expressing Ube2o shRNA together with Prkaa2 shRNA were subjected to immunoblotting for the indicated proteins. (J) Insulin-stimulated glucose uptake rate of C2C12 myotubes from I. n = 3. Error bars represent ±SEM. P value was determined by ANOVA (***P < 0.001, ###P < 0.001).

Journal: JCI Insight

Article Title: A muscle-specific UBE2O/AMPK α 2 axis promotes insulin resistance and metabolic syndrome in obesity

doi: 10.1172/jci.insight.128269

Figure Lengend Snippet: (A) UBE2O (top) or AMPKα2 (bottom) immunoprecipitates of lysates from skeletal muscle of control (Ube2ofl/fl) and Ube2oΔMus mice were subjected to immunoblotting for AMPKα2 or UBE2O. (B) Lysates from C2C12 myotubes transfected with the indicated plasmids were subjected to metal affinity purification for His-tagged ubiquitin (His-Ub), then immunoblotting for ubiquitinated AMPKα2. Ni-NTA, Ni2+-nitrilotriacetic acid. (C) Recombinant AMPKα2 proteins were subjected to in vitro ubiquitination assay in the presence of in vitro translated WT or C1037S (CS) mutant UBE2O. (D) Lysates from C2C12 myotubes expressing WT or CS mutant UBE2O treated with MG132 (10 μM) for 6 hours were subjected to immunoblotting. (E) Lysates from C2C12 myotubes expressing WT or CS mutant UBE2O treated with cycloheximide (CHX) for the indicated times were subjected to immunoblotting (top). AMPKα2 or AMPKα1 protein levels were quantified by normalizing to the intensity of the HSP90 band (bottom). (F) Lysates from skeletal muscle of 12-week-old Ube2o+/+, Ube2o–/–, Ube2ofl/fl, and Ube2oΔMus mice were subjected to immunoblotting for the indicated proteins. (G) Lysates from C2C12 myotubes expressing UBE2O shRNA together with exogenous WT or CS mutant UBE2O from a Ube2o ORF transcript lacking the 3′ UTR sequence targeted by shRNA (shUbe2o-3′UTR) were subjected to immunoblotting for the indicated proteins. (H) Insulin-stimulated glucose uptake rate of C2C12 myotubes from G. 2-DG6P, 2-deoxyglucose-6-phosphate. n = 4. (I) Lysates from C2C12 myotubes expressing Ube2o shRNA together with Prkaa2 shRNA were subjected to immunoblotting for the indicated proteins. (J) Insulin-stimulated glucose uptake rate of C2C12 myotubes from I. n = 3. Error bars represent ±SEM. P value was determined by ANOVA (***P < 0.001, ###P < 0.001).

Article Snippet: The mutant construct (C1037S) of Flag-tagged mouse UBE2O (Origene) was generated by site-directed mutagenesis and cloned into pBabe retroviral vector.

Techniques: Western Blot, Transfection, Affinity Purification, Recombinant, In Vitro, Ubiquitin Assay, Mutagenesis, Expressing, shRNA, Sequencing