Journal: PLoS ONE

Article Title: Pharmacological Blockade of Cannabinoid CB 1 Receptors in Diet-Induced Obesity Regulates Mitochondrial Dihydrolipoamide Dehydrogenase in Muscle

doi: 10.1371/journal.pone.0145244

Figure Lengend Snippet: Primer References for TaqMan® Gene Expression Assays (ThermoFisher).

Article Snippet: Eno3 , Rn01464911_m1 , 77.

Techniques: Gene Expression, Amplification

Journal: PLoS ONE

Article Title: Pharmacological Blockade of Cannabinoid CB 1 Receptors in Diet-Induced Obesity Regulates Mitochondrial Dihydrolipoamide Dehydrogenase in Muscle

doi: 10.1371/journal.pone.0145244

Figure Lengend Snippet: Proteins Identified using 2D Electrophoresis and MALDI-TOF/LC-ESI MS Analyses.

Article Snippet: Eno3 , Rn01464911_m1 , 77.

Techniques: Two-Dimensional Gel Electrophoresis

Journal: PLoS ONE

Article Title: Pharmacological Blockade of Cannabinoid CB 1 Receptors in Diet-Induced Obesity Regulates Mitochondrial Dihydrolipoamide Dehydrogenase in Muscle

doi: 10.1371/journal.pone.0145244

Figure Lengend Snippet: AM251 effects on the protein expressions of GPI, TPI, Eno3, PKM1, LDHa and Glo1 in the abdominal muscle of STD, HFD and HCD-fed rats ( A-F ). Representative 2D polyacrylamide gels showing the intensity of the identified spots ( G ). Bonferroni’s test ( n = 6): * P <0.05, ** P <0.01, *** P <0.001 vs . STD-vehicle; # P <0.05, ## P <0.01, ### P <0.001 vs . HFD-vehicle; $ P <0.05, $ $ $ P <0.001 vs . HCD-vehicle.

Article Snippet: Eno3 , Rn01464911_m1 , 77.

Techniques:

Journal: PLoS ONE

Article Title: Pharmacological Blockade of Cannabinoid CB 1 Receptors in Diet-Induced Obesity Regulates Mitochondrial Dihydrolipoamide Dehydrogenase in Muscle

doi: 10.1371/journal.pone.0145244

Figure Lengend Snippet: AM251 effects on the gene expressions of Gpi , Tpi1 , Eno3 , Pkm , Ldha and Glo1 in the abdominal muscle of STD, HFD and HCD-fed rats. Bonferroni’s test ( n = 8): * P <0.05, *** P <0.001 vs . STD-vehicle; $ P <0.05 vs . HCD-vehicle.

Article Snippet: Eno3 , Rn01464911_m1 , 77.

Techniques:

Journal: PLoS ONE

Article Title: Pharmacological Blockade of Cannabinoid CB 1 Receptors in Diet-Induced Obesity Regulates Mitochondrial Dihydrolipoamide Dehydrogenase in Muscle

doi: 10.1371/journal.pone.0145244

Figure Lengend Snippet: Gene ( A ) and protein ( B ) expressions of GPI, TPI, Eno3, PKM1, LDHa and Glo1 in the abdominal muscle of wild-type and CB 1 -/- mice. C) Representative immunoblots. Student’s t test ( n = 6): * P <0.05 vs . WT.

Article Snippet: Eno3 , Rn01464911_m1 , 77.

Techniques: Western Blot

Journal: Cell reports

Article Title: A Dimerization Function in the Intrinsically Disordered N-Terminal Region of Src

doi: 10.1016/j.celrep.2018.09.035

Figure Lengend Snippet: (A and B) SYF cells were transfected with the indicated constructs and cell lysates immunoblotted as indicated. (C) The indicated full-length or partial length Src constructs were expressed and purified from dasatinib-treated HEK293T cells. Dasatinib treatment was to enable the purification of unphosphorylated Src. Coomassie stains of the purified proteins are shown here. (D) In vitro kinase reactions were carried with 10 nM of the indicated purified V5-tagged Src proteins for 10 min and assayed by anti-pY419Src immunoblotting. ATP was omitted in the negative control arms. (E) The same in vitro kinase reactions were performed using different concentrations of protein as indicated. The quantified autophosphorylation results are shown here, and the corresponding immunoblots of these reactions are shown in . The data points represent the average of n = 2; errors bars represent SEM. (F) Three separate in vitro kinase reactions were performed using 4 nM Src proteins and each of the three indicated purified recombinant substrates. Kinase reaction linear conditions were previously established and are as follows: 40 nM paxillin in reaction for 20 min, 250 nM enolase in reaction for 20 min, and 70 nM Trask/CDCP1 in reaction for 10 min. Substrate phosphorylation was assayed by anti-pTyr immunoblotting and kinase and substrate proteins immunoblotted with V5 and substrate-specific antibodies. ATP was omitted in the negative control arms. (G) The indicated cell lines were treated with 1 µM DDD85646 for 30 hr and cell lysates immunoblotted as indicated. Anti-pY416Src antibodies cross-react with all members of the Src family. (H) The same lysates were assayed specifically for Src autophosphorylation by anti-Src immunoprecipitation and immunoblotted as indicated. Experiments with the various constructs were performed 2–3 times, and representative data are shown here.

Article Snippet: Recombinant human Paxillin was purchased from Raybiotech (Norcross, GA) and recombinant Enolase (ENO3) was purchased from Sino Biological Inc.(Beijing, PRC).

Techniques: Transfection, Construct, Purification, In Vitro, Western Blot, Negative Control, Recombinant, Immunoprecipitation

Journal: Cell reports

Article Title: A Dimerization Function in the Intrinsically Disordered N-Terminal Region of Src

doi: 10.1016/j.celrep.2018.09.035

Figure Lengend Snippet:

Article Snippet: Recombinant human Paxillin was purchased from Raybiotech (Norcross, GA) and recombinant Enolase (ENO3) was purchased from Sino Biological Inc.(Beijing, PRC).

Techniques: Control, Virus, Recombinant, Kinase Assay, Membrane, Protein Extraction, Modification, Software

Journal: bioRxiv

Article Title: Proteogenomic single cell analysis of skeletal muscle myocytes

doi: 10.1101/2020.01.23.916791

Figure Lengend Snippet: a) HPA images (top row) are mosaic for DCAF11 and ENO3 and negative for PVALB staining. Follow up staining validated the DCAF11 and ENO3 staining while suggesting a subtle mosaicism of PVALB in humans. In mice, ENO3 and PVALB are clearly mosaic, while DCAF11 is not. b) RNA-ISH demonstrates co-expression of CYB5R1 and ENO3 in a mosaic pattern. c) Only Eno3 was observed (in a mosaic pattern) in mouse muscle by RNA-ISH.

Article Snippet: Antibodies were obtained for WDR23/DCAF11 (bs-8388R, Bioss Antibodies), PVALB (A2781, Abclonal), and ENO3 (ARP48203_T100, Aviva Systems Biology) that were reported to cross react to both human and mouse.

Techniques: Staining, Expressing

Journal: Nutrients

Article Title: MicroRNA-34a Mediates High-Fat-Induced Hepatic Insulin Resistance by Targeting ENO3

doi: 10.3390/nu15214616

Figure Lengend Snippet: ENO3 is specifically repressed by miR-34a as a direct target gene. ( a ) Sequence alignment between miR-34a and the 3′UTR of ENO3. ( b ) The effect of miR-34a on the 3′UTR of ENO3 was determined by luciferase activity assays. HEK-293 cells were co-transfected with constructed WT or MUT 3′UTR of ENO3 plasmid, miR-34a and its inhibitor for 48 h, and luciferase activities were measured. ( c ) The changes in the expression level of ENO3 protein in AML-12 cells with different treatments. The AML-12 cells were transfected with 100 nM miR-34a mimic or 200 nM miR-34a inhibitor for 24 h and then treated with or without 400 μM PA for 24 h. A scrambled sequence was used as a negative control (NC). ( d , e ) The changes in the expression level of ENO3 protein detected by Western blot ( d ) and immunohistochemical staining (scale bars, 100 μM) ( e ) in the liver of mice of different groups. After 12 weeks of feeding with ND or HFD, the mice were administered AAV8-mediated liver-specific miR-34a mimic or inhibitor by tail vein, respectively. Then, the mice continued to be fed an ND or HFD for 6 weeks. * p < 0.05 and ** p < 0.01 for the indicated comparison, or vs. NC or ND; †† p < 0.01 vs. PA or HFD.

Article Snippet: To determine the role of miR-34a and ENO3 in high-fat-induced hepatic IR in vitro, miR-34a mimic, miR-34a inhibitor (GenePharma, Shanghai, China), ENO3 siRNA and ENO3 overexpression plasmid (RiboBio, Guangzhou, China) were applied in cell experiments.

Techniques: Sequencing, Luciferase, Activity Assay, Transfection, Construct, Plasmid Preparation, Expressing, Negative Control, Western Blot, Immunohistochemistry, Staining, Comparison

Journal: Nutrients

Article Title: MicroRNA-34a Mediates High-Fat-Induced Hepatic Insulin Resistance by Targeting ENO3

doi: 10.3390/nu15214616

Figure Lengend Snippet: ENO3 participates in miR-34a-mediated hepatic IR in AML12 cells. ( a ) The changes in the expression levels of ENO3, p-IRS2 (Ser1100) and p-Akt (Ser473) proteins in AML-12 cells transfected with ENO3 siRNAs. The AML12 cells were transfected 100 nM three pairs of siRNA targeted to ENO3 gene (siRNA-1, siRNA-2 and siRNA-3, respectively) for 48 h, and related proteins were measured. ( b ) The changes in the expression levels of ENO3, p-IRS2 (Ser1100) and p-Akt (Ser473) proteins in AML12 cells transfected with ENO3 overexpression plasmid. The AML12 cells were transfected 1 μg/mL ENO3 overexpression plasmid for 48 h, and related proteins were detected. ( c ) The changes in the expression levels of ENO3, p-IRS2 (Ser1100) and p-Akt (Ser473) proteins in AML12 cells treated with miR-34a or/and ENO3 overexpression plasmid. The AML12 cells were transfected with 100 nM miR-34a mimic or/and 1 μg/mL ENO3 overexpression plasmid for 48 h, and related proteins were tested. The scrambled sequence was used as a negative control (NC). * p < 0.05 and ** p < 0.01 vs. NC; † p < 0.05 and †† p < 0.01 vs. miR-34a.

Article Snippet: To determine the role of miR-34a and ENO3 in high-fat-induced hepatic IR in vitro, miR-34a mimic, miR-34a inhibitor (GenePharma, Shanghai, China), ENO3 siRNA and ENO3 overexpression plasmid (RiboBio, Guangzhou, China) were applied in cell experiments.

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, Sequencing, Negative Control

Journal: Nutrients

Article Title: MicroRNA-34a Mediates High-Fat-Induced Hepatic Insulin Resistance by Targeting ENO3

doi: 10.3390/nu15214616

Figure Lengend Snippet: ENO3 is involved in PA-induced hepatic IR in AML12 cells. ( a – e ) The changes in the expression levels of ENO3, p-IRS2 (Ser1100) and p-Akt (Ser473) ( a ), glucose uptake (scale bars, 100 μM) ( b ), glucose consumption ( c ), glycogen content ( d ) and glucose production ( e ) in AML-12 cells with different treatments. The AML-12 cells were transfected with 100 nM scrambled sequence, 1 μg/mL ENO3 overexpression plasmid for 24 h and then treated with or without 400 μM PA for 24 h. A scrambled sequence was used as a negative control (NC). * p < 0.05 and ** p < 0.01 vs. NC; †† p < 0.01 vs. PA.

Article Snippet: To determine the role of miR-34a and ENO3 in high-fat-induced hepatic IR in vitro, miR-34a mimic, miR-34a inhibitor (GenePharma, Shanghai, China), ENO3 siRNA and ENO3 overexpression plasmid (RiboBio, Guangzhou, China) were applied in cell experiments.

Techniques: Expressing, Transfection, Sequencing, Over Expression, Plasmid Preparation, Negative Control

Journal: Nutrients

Article Title: MicroRNA-34a Mediates High-Fat-Induced Hepatic Insulin Resistance by Targeting ENO3

doi: 10.3390/nu15214616

Figure Lengend Snippet: ENO3 is involved in HFD-induced hepatic IR in mice. ( a – e ) The changes in blood glucose during IPGTT ( a ), the AUC of glucose between 0 and 120 min of IPGTT ( b ), serum insulin levels ( c ), HOMA-IR values ( d ), the expression levels of ENO3, p-IRS2 (Ser1100), p-Akt (Ser473) proteins ( e ) in the liver of mice of different groups. After 11 weeks of being fed with ND or HFD, the mice were administered AAV8-mediated liver-specific ENO3 expression plasmid or scrambled sequence via the tail vein, respectively. Then, the mice continued to be feed an ND or HFD for 8 weeks. * p < 0.05 and ** p < 0.01 vs. ND; † p < 0.05 and †† p < 0.01 vs. HFD.

Article Snippet: To determine the role of miR-34a and ENO3 in high-fat-induced hepatic IR in vitro, miR-34a mimic, miR-34a inhibitor (GenePharma, Shanghai, China), ENO3 siRNA and ENO3 overexpression plasmid (RiboBio, Guangzhou, China) were applied in cell experiments.

Techniques: Expressing, Plasmid Preparation, Sequencing

Journal: Nutrients

Article Title: MicroRNA-34a Mediates High-Fat-Induced Hepatic Insulin Resistance by Targeting ENO3

doi: 10.3390/nu15214616

Figure Lengend Snippet: Validation of changes in miR-34a/ENO3 pathway in the liver of patients with NAFLD. The changes in serum levels of glucose ( a ), insulin ( b ), HOMA-IR values ( c ), the level of miR-34a ( d ), the expression levels of ENO3, p-IRS2 (Ser1100) and p-Akt (Ser473) proteins detected by Western blot ( e ) and immunohistochemical staining of ENO3 and p-Akt proteins (scale bars, 100 μM) ( f ) in the liver of control subjects and patients with NAFLD. ** p < 0.01 vs. Control.

Article Snippet: To determine the role of miR-34a and ENO3 in high-fat-induced hepatic IR in vitro, miR-34a mimic, miR-34a inhibitor (GenePharma, Shanghai, China), ENO3 siRNA and ENO3 overexpression plasmid (RiboBio, Guangzhou, China) were applied in cell experiments.

Techniques: Expressing, Western Blot, Immunohistochemistry, Staining