1 1 dihexadecyl 3 3 3 3 tetramethylindocarbocyanine perchlorate diic16 3

Plasmids expressing de novo purine biosynthetic enzymes conjugated with monomeric enhanced green fluorescent protein (EGFP) or monomeric orange fluorescent protein (OFP) were mostly prepared previously.^{1 (link)} Note that EGFP in this article contains three point mutations, A206K, L221K, and F223R, which prevent potential GFP-mediated oligomerization of tagged proteins.^{36 (link)} We also prepared a new plasmid expressing EGFP-ATIC using the pEGFP-C1 vector (Clontech) to replace the previous clone of GFP-ATIC,^{1 (link)} which was constructed to express GFP_S65T-fusion proteins from the pC1-Neo vector (Promega). The cloned plasmid in this work was confirmed by restriction enzyme digestions and DNA sequencing (GeneWiz). The plasmids expressing shRNA_AMPK (cat# sc-45312-SH) and shRNAControl (cat# sc-108060) were purchased from Santa Cruz.
5-Aminoimidazole-4-carboxamide ribonucleoside (AICAR) and ribonucleotide (ZMP) were obtained from EMD Biosciences. Metformin, phenformin, IMP, AMP, and GMP were obtained from Sigma-Aldrich. A769662 was obtained from Santa Cruz Biotechnology. ATP was obtained from Invitrogen. Cellular staining reagents, including LysoTracker Red DND-99 (cat# L7528), 1,1′-dihexadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiIC₁₆(3); cat# D384), and MitroTrackerOrange CMTMRos (cat# M7510), were obtained from Molecular Probes (Life Technologies).