Supplementary Materials and Methods from Early Adaptation and Acquired Resistance to CDK4/6 Inhibition in Estrogen Receptor–Positive Breast Cancer
Year: 2023
Type: supplementary-materials
Abstract: Supplementary Materials and Methods Antibodies and reagentsAntibodies used were phospho-AKT Ser473 (4058), phospho-AKT T308 ( 2965), AKT (4691), phospho-ERK1/2-Thr202/Tyr204 (4370), ERK1/2 (9102), phospho-pRb S807/811 (8516), phospho-pRb Ser 780 (9307), pRb (9313), cyclin D1 (2978), cyclin E2 (4132), cyclin E1 (4129), CDK2 (2546), CDK4 (12790), S6 (2317), phospho-S6 Ser235/6 (2111), phospho-S6 Ser240/4 (2215), PARP cleavage (5625 and 9441) (all Cell Signalling Technology, Danvers, MA), β-actin (A5441) and Tubulin (T-9026)(Sigma), CDK2 (sc163), Cyclin E1 (sc198), Human GAPDH (sc137179) (Santa Cruz Biotechnology), Cyclin E2 (ab40890) (AbCAM), Cyclin D1 (RM9104) (Thermo-Scientific).PD-0332991 (palbociclib) was provided by Pfizer.GDC-0941 and fulvestrant were purchased from Selleckchem.NVP-AEW541 from Cayman Chemicals.siRNA were from Dharmacon.Unless otherwise stated palbociclib was used at 500nM and GDC-0941 at 250nM. Western BlottingCell lines were grown on 35mm plates, treated as indicated, and lysed in NP40 lysis buffer, (1% v/v NP40, 10mM Tris.Cl pH8, 150mM NaCl, 1mM EDTA, 1mM DTT) supplemented with phosphatase inhibitor cocktail (Cell Signalling Technology, 5872)
Authors: Maria Teresa Herrera-Abreu, Marta Palafox, Uzma Asghar, Martín A. Rivas, Rosalind J. Cutts, Isaac Garcia-Murillas, Alex Pearson, Marta Guzman, Olga Rodriguez, Judit Grueso, Meritxell Bellet, Javier Cortés, Richard Elliott, Sunil Pancholi, Christopher J. Lord, José Baselga, Mitch Dowsett, Lesley-Ann Martin, Nicholas C. Turner, Violeta Serra
Language: English
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4361876490