Synthesis and Characterization of HI-511 and APIO-EE-09

HI-511 and APIO-EE-09 31 (link) were synthesized and the structure of HI-511 was confirmed by NMR spectroscopy. Tris, NaCl, and SDS for molecular biology and buffer preparation were purchased from Sigma-Aldrich (St. Louis, MO). All cell culture media were purchased from Invitrogen (Waltham, MA). Antibodies to detect cleaved PARP (#5625), cleaved caspase-3 (#9664), p-histone H3 (#9701), histone H3 (#9715), p-BRAF (#2696), p-MEK (#9121), MEK (9122), p-ERKs (#9101), ERKs (#9102), p-PI3-K (#4228), PI3-K (#3011), p-AKT (#9271), AKT (#9272), PCNA (D3H8P; #13110) were obtained from Cell Signaling Technology (Danvers, MA). β-Actin (sc-47778), PARP-1 (sc-74470), caspase-3 (sc-7272), Bcl-2 (sc-7382), Bax (sc-20067), and GAPDH (sc-32233) antibodies were from Santa Cruz Biotechnology (Dallas, TX). BRAF (07-583) was from MilliporeSigma (Burlington, MA), AURKB (NB100-294) was from Novus Biologicals (Centennial CO), and vemurafenib was purchased from Cayman Chemical Co. (Ann Arbor, MI).

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Chang X., Zhang T., Wang Q., Rathore M.G., Reddy K., Chen H., Shin S.H., Ma W.Y., Bode A.M, & Dong Z. (2020). HI-511 overcomes melanoma drug resistance via targeting AURKB and BRAF V600E. Theranostics, 10(21), 9721-9740.

Publication 2020

NaCl cleaved PARP cleaved caspase-3 p-histone H3 histone H3 p-BRAF p-MEK p-PI3-K p-AKT GAPDH (sc-32233) vemurafenib

Actin Antibodies Bcl 2 Biologicals Braf Buffer Caspase 3 Cayman Cell Cell culture Culture media Erks Gapdh Histone h3 Nacl Nmr spectroscopy Novus Parp 1 Pcna Tris Vemurafenib

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Variable analysis

independent variables

HI-511
APIO-EE-09 31

dependent variables

Structure of HI-511 confirmed by NMR spectroscopy

control variables

Cell culture media

controls

Positive controls: None specified
Negative controls: None specified

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