Additional file 1 of First in class dual MDM2/MDMX inhibitor ALRN-6924 enhances antitumor efficacy of chemotherapy in TP53 wild-type hormone receptor-positive breast cancer models
Creators: name: Pairawan, Seyed
name: Zhao, Ming
name: Yuca, Erkan
name: Annis, Allen
name: Evans, Kurt
name: Sutton, David
name: Carvajal, Luis
name: Ren, Jian-Guo
name: Santiago, Solimar
name: Guerlavais, Vincent
name: Akcakanat, Argun
name: Tapia, Coya
name: Yang, Fei
name: Bose, Priya Subash Chandra
name: Zheng, Xiaofeng
name: Dumbrava, Ecaterina Ileana
name: Aivado, Manuel
name: Meric-Bernstam, Funda
Publisher: name: figshare
Year: 2021
Subjects: Biochemistry
Space Science
Medicine
Cell Biology
Molecular Biology
Pharmacology
Biological Sciences not elsewhere classified
Cancer
Computational Biology

description: Additional file 1: Supplementary Figure 1. ALRN-6924 combined with eribulin synergistically inhibit clonogenic capacity of MCF-7 breast cancer cell line. MCF-7 colony formation assay treated with either vehicle, ALRN-6924, paclitaxel, or combination of both. Total colony area was measured by using NIH ImageJ software. Mean +/- SD is demonstrated. Supplementary Figure 2. In vitro validation of in vivo functional proteomic changes after treatment with ALRN-6924 and paclitaxel. A. MCF-7 cells were treated with vehicle, ALRN-6924 or paclitaxel in a dose-dependent fashion for 24 hours followed by immunoblotting with indicated antibodies. B. Quantification of relative p53 levels by standardizing to β-actin. C. Quantification of relative p21 levels by standardizing to β-actin. D. Quantification of relative c-Myc levels by standardizing to β-actin. E. Quantification of relative p-Rb (Ser807/811) levels by standardizing to β-actin. F. Quantification of relative LC3-I/LC3-II ratio levels by standardizing to β-actin. G. Quantification of relative LC3 levels by standardizing to β-actin. H. Quantification of relative p-S6 (Ser235/236) levels by standardizing to β-actin. Supplementary Figure 3. Immunoblotting of signaling pathways affected by ALRN-6924 combined with paclitaxel. A. MCF-7 cells were treated with vehicle, ALRN-6924, paclitaxel or combination of both in a dose-dependent fashion for 24 hours followed by immunoblotting with indicated antibodies. B. Quantification of relative p53 levels by standardizing to β-actin. C. Quantification of relative p21 levels by standardizing to β-actin. D. Quantification of relative p-Rb (Ser807/811) levels by standardizing to β-actin. E. Quantification of relative LC3-I/LC3-II ratio levels by standardizing to β-actin. Supplementary Figure 4. MDMX knockout cell line models. A. Immunoblotting of MDMX. Two MDMX knockout clones (M1 and M4, equally KO-1 and KO-2) were established in Myeloid leukemia cell line OCI-AML3 cells. MDMX was blotted with anti-MDMX antibody. B. Cell proliferation assay. Cell proliferation of two MDMX knockout cell line clones and control cell line was examined using the same methods as described in Fig. 1. C-D. Drug sensitivity assay. Two MDMX knockout cell line clones and control cell line were treated ALRN-6924 (C) and RG7388 (D) to assess the efficacy of these drugs using the same method as described in Fig. 1. Supplementary Table 1. Cell line panels for drug screening. Supplementary Table 2. IPA analysis of MCF-7 ALRN-6924 + Paclitaxel vs Control.