Supplementary Figure S2 from MEK Inhibitor Selumetinib (AZD6244; ARRY-142886) Prevents Lung Metastasis in a Triple-Negative Breast Cancer Xenograft Model
Year: 2023
Type: supplementary-materials
Abstract: MDA-MB-231 cells were transfected with siMEK1/2 or scrambled control siRNA. 48 hours after transfection cells were plated in 3D culture and treated with selumetinib for 48 hours. For western blot analysis, cell pellets were lysed as described previously with β-actin used as a loading control (16). Primary antibodies were anti-phospho-p42/44 MAPK (Thr202/Tyr204) (1:1000 dilution; Cell Signaling), anti-β-actin (1:5000 dilution; Sigma-Aldrich), anti-fibronectin (1:500 dilution; BD Transduction), anti-vimentin (1:1000 dilution; Cell Signaling), anti-E-cadherin (1:1000 dilution; BD Transduction), anti-beta-catenin (1:1000 dilution; Cell Signaling). Secondary antibodies were horseradish peroxidase-conjugated IgG (1:10,000 dilution; Invitrogen) for chemiluminescent signal detection and the corresponding Alexa Fluor-conjugated IgG (1:5000 dilution; Invitrogen) for fluorescence signal detection.
Language: English
Topic: Melanoma and MAPK Pathways
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4362484703