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3 protocols using dimethyl succinate

1

Metabolic Regulation Pathway Investigation

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3-mercaptopicolinic acid, Z-VAD-FMK and Necrostatin-1 were purchased from Santa Cruz Biotechnology. Dimethyl-2-oxoglutarate, dimethyl-fumarate, dimethyl-succinate and dimethyl-malate were purchased from Tokyo Chemical Industry, Sodium pyruvate and L-glutamine were purchased from Thermo Fisher Scientific, N-acetylcysteine (NAC) was purchased from Sigma-Aldrich, and dichlorofluorescin diacetate from Beyotime.
Antibodies against PCK1 (ab28455, Abcam), PCK2 (ab70359, Abcam) and YAP1 (ab52771, Abcam), phospho-AMPKα (Thr172) (Cell Signaling Technology, 2535), AMPKα (Cell Signaling, 2532), phospho-ACC (Ser79) (Cell Signaling, 11818) and phospho-c-Jun (Ser73) (Cell Signaling, 3270) were purchased commercially.
pBABE-Flag-PCK1 and pQCXIH-Flag-PCK2 were cloned from cDNA provided by Dr. Jiahuai Han (Xiamen University, Xiamen, China). The PB[CMV-myc-YAP-5SA]DS, PB[Act-RFP]DS and Act-PB Transposase plasmids were generous gifts from Dr. Bin Zhao (Zhejiang University, Hangzhou, China). The PB[CMV-flag-PCK1]DS donor plasmids were constructed by excising Act-RFP from the PB[Act-RFP]DS plasmid and ligating the corresponding fragments excised from pQCXIH vectors.
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2

Metabolic Regulation Pathway Investigation

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3-mercaptopicolinic acid, Z-VAD-FMK and Necrostatin-1 were purchased from Santa Cruz Biotechnology. Dimethyl-2-oxoglutarate, dimethyl-fumarate, dimethyl-succinate and dimethyl-malate were purchased from Tokyo Chemical Industry, Sodium pyruvate and L-glutamine were purchased from Thermo Fisher Scientific, N-acetylcysteine (NAC) was purchased from Sigma-Aldrich, and dichlorofluorescin diacetate from Beyotime.
Antibodies against PCK1 (ab28455, Abcam), PCK2 (ab70359, Abcam) and YAP1 (ab52771, Abcam), phospho-AMPKα (Thr172) (Cell Signaling Technology, 2535), AMPKα (Cell Signaling, 2532), phospho-ACC (Ser79) (Cell Signaling, 11818) and phospho-c-Jun (Ser73) (Cell Signaling, 3270) were purchased commercially.
pBABE-Flag-PCK1 and pQCXIH-Flag-PCK2 were cloned from cDNA provided by Dr. Jiahuai Han (Xiamen University, Xiamen, China). The PB[CMV-myc-YAP-5SA]DS, PB[Act-RFP]DS and Act-PB Transposase plasmids were generous gifts from Dr. Bin Zhao (Zhejiang University, Hangzhou, China). The PB[CMV-flag-PCK1]DS donor plasmids were constructed by excising Act-RFP from the PB[Act-RFP]DS plasmid and ligating the corresponding fragments excised from pQCXIH vectors.
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3

Organocatalytic Ring-Opening Polymerization

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All manipulations involving air- and/or water-sensitive compounds were carried out with the standard Schlenk and vacuum line techniques under an argon atmosphere or in a nitrogen-filled glovebox. Succinic anhydride (SA) and citraconic anhydride (CA) were purchased from Macklin. Propylene epoxide (PO) was purchased from Aldrich. Chromatographic purity solvents, including toluene, dichloromethane, tetrahydrofuran (THF), ethanol, and flaky sodium hydroxide (CaH2), were bought from SINOPHARM. Triethyl borane (TEB) in tetrahydrofuran solution (1.0 mol/L), Dimethyl succinate (DS), and dimethyl citraconate (DC) were purchased from the Tokyo Chemical Industry (TCI). Triethylamine (TEA) was purchased from Aldrich, and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), 7-methyl-1,5,7-triazabicyclo[4.4.0]dec-5-ene (mTBD) and 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD) were purchased from TCI. PO, TEA, DBU, DS, DC, and toluene were refluxed over CaH2 for 24 h and vacuum-distilled prior to use. In particular, chloroform-d for dynamic NMR characterization of amine-ester complexes was refluxed over CaH2 for 24 h and distilled prior to use. Both the anhydrides (SA and CA) were purified by vacuum sublimation three times. The purified anhydrides were collected under an inert atmosphere and stored in the glovebox for use. Other organic reagents were used without purification.
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