Rg7388

ALRN-6924 was kindly provided by Aileron Therapeutics Inc. (Watertown, MA, USA). RG7112 and RG7388 (idasanutlin) and paclitaxel were purchased for in vitro studies from MedChemExpress (Monmouth Junction, NJ, USA). For in vivo paclitaxel efficacy studies, the agent was obtained from MD Anderson Cancer Center pharmacy. This source of paclitaxel was later changed to the National Cancer Institute for pharmacodynamic experiments. paclitaxel was prepared in 15% ethanol, 15% kolliphor (Sigma-Aldrich Co., LLC, St Louis, MO, USA), and water for in vivo studies. Eribulin was obtained from the MD Anderson Cancer Center pharmacy. Dimethyl sulfoxide (DMSO) was obtained from Sigma-Aldrich (Sigma-Aldrich Co., LLC, St Louis, MO, USA). All drugs used for in vitro studies were initially prepared in DMSO prior to dilution in a tissue culture medium.

Human AML cell lines, including MV-4-11, MOLM-13, and HL-60, were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). Human AML cell line Ontario Cancer Institute‒Acute Myeloid Leukemia-3 (OCI-AML-3) was sourced from Cobioer (Nanjing, China) and SKM-1 from the Japanese Collection of Research Bioresources Cell Bank (JCRB). MV-4–11 cells were propagated in Iscove’s Modified Dulbecco’s Medium (IMDM, Gibco, Grand Island, NY, USA Cat# C12440500BT) supplemented with 10% fetal calf serum (FCS, AUSGENEX, Loganholme, QLD, Australia, Cat# FBSSA500-S). Other cell lines were propagated in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, Cat# C11875500BT) containing 10% FCS. All experiments utilized genetically authenticated, microbial-free cells in their exponential phases of growth.
APG-115 (produced by Ascentage Pharma, Jiangsu, China) was dissolved in dimethyl sulfoxide (DMSO, Sigma, St. Louis, MO, USA, Cat# D8418) for in vitro experiments or suspended in 0.2% hydroxypropyl methylcellulose (HPMC, Sigma, Cat# H7509-25G) for in vivo animal studies. DAC (Cat# S120009), AZA (Cat# S178206), and Ara-C (Cat# S164806) purchased from Selleckchem were dissolved in DMSO for in vitro assays and saline for in vivo studies. RG-7388 (idasanutlin, MDM2-P53 inhibitor, MedChemExpress, Cat# HY-15676) was dissolved in DMSO for in vitro assays.

The histone deacetylase (HDAC) inhibitor SAHA was purchased from Selleckchem (Houston, TX, USA), the MDM2 inhibitor RG7388 was purchased from MedChemExpress (Monmouth Junction, NJ, USA). Letrozole was purchased from Selleckchem (Houston, TX, USA), and Necrostatin was purchased from Abcam (Cambridge, MA, USA). The primary antibodies against p53, phsoph-p53, p21, CDC25C, TIPM-1, CDK1, BAK, BAX, APAF1, Bcl-XL, RIP1, and cleaved PARP (1:1000) were purchased from Cell Signaling Technology (Danvers, MA, USA). Phospho-RIP3 and MLKL (1:1000) antibodies were purchase from Abcam (Cambridge). MDM2 (1:500) was purchased from Santa Cruz biotechnology (Dallas, TX, USA). The β-actin antibody (1:2000) was purchased from Sigma Aldrich chemical company (St. Louis, MO, USA). The secondary antibodies anti-rabbit, anti-mouse, HRP conjugated, and DMSO were purchased from Sigma Aldrich chemical company. Nitrocellulose membranes (0.45 µm) were purchased from Amersham (GE Healthcare Life Sciences, Marlborough, MA, USA). ECL was purchased from KPL biosolutions (Milford, MA, USA). SYTOX^® Green and DEVD-amc CellEvent^TM Caspase-3/7 Green ReadyProbe^TM were purchased from Thermo Fisher (Molecular Probes, Life Technologies, Carlsbad, CA, USA). All other chemicals used in this study were of research grade.