Fluorescent PFK1 Fusion Protein Protocol

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).

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Schmitt D.L., Dranchak P., Parajuli P., Blivis D., Voss T., Kohnhorst C.L., Kyoung M., Inglese J, & An S. (2023). High-throughput screening identifies cell cycle-associated signaling cascades that regulate a multienzyme glucosome assembly in human cells. PLOS ONE, 18(8), e0289707.

Publication 2023

DMSO resveratrol clemastine fumarate ARP101 1,10-phenanthroline kenpaullone calmidazolium chloride PAC-1 SU9516 SL-0101-1 palbociclib hesperadin

Bisindolylmaleimide ii Bs 181 Calmidazolium chloride Cayman Cells Clemastine fumarate Dmso Enhanced green fluorescent protein Hesperadin Kenpaullone Palbociclib Pf 03814735 Phenanthroline Plasmid Point mutations Resveratrol Su9516 Triflupromazine

Corresponding Organization : National Human Genome Research Institute

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Variable analysis

independent variables

Small molecules used in this study, including resveratrol, clemastine fumarate, ARP101, 1,10-phenanthroline, triflupromazine hydrochloride, kenpaullone, calmidazolium chloride, PAC-1, SU9516, SL-0101-1, palbociclib, BS-181, PF-03814735, hesperadin, bisindolylmaleimide II (BIM II), and BIM X

dependent variables

Not explicitly mentioned

control variables

DMSO (Sigma) was used as the solvent for the small molecules

controls

No positive or negative controls were explicitly mentioned in the information provided.

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