Pac 1

A plasmid expressing PFK1 conjugated with a monomeric enhanced green fluorescent protein (mEGFP) was prepared previously [4 (link)]. Note that mEGFP contains three point mutations, A206K, L221K, and F223R, which prevent oligomerization of EGFP in cells [30 (link),31 (link)]. Small molecules used in this study were dissolved in DMSO (Sigma) and used at the concentrations indicated, that include resveratrol (Sigma), clemastine fumarate (Sigma), ARP101 (Tocris), 1,10-phenanthroline (Acros Organics), triflupromazine hydrochloride (Sigma), kenpaullone (Sigma), calmidazolium chloride (Sigma), PAC-1 (Sigma), SU9516 (Tocris), SL-0101-1 (Tocris), palbociclib (Sigma), BS-181 (Tocris), PF-03814735 (Sigma or Cayman), hesperadin (Tocris), bisindolylmaleimide II (BIM II, Cayman), and BIM X (Cayman).

PAC-1, BP (2,2′-dipyridyl, a well-known iron (II) chelator and chemical hypoxia mimic), MG132, and ferrozine were purchased from Sigma-Aldrich (St. Louis, MO). Other reagents and all cell culture media were purchased from Thermo Fisher unless specifically stated. Primary antibodies targeting pH2AX (cat. no. H10292) and BrdU and labeled secondary antibodies were obtained from Life Technologies (Beverly, MA). Primary antibodies targeting hypoxia-inducible factor 1α (HIF1α; cat. no. ab16066) and Pro564 hydroxylated HIF1α (cat. no. 3434) were purchased from Abcam (Cambridge, MA) and Cell Signaling Technology (Danvers, MA), respectively.