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.
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.