2024-04-22
ENA first public
ERC000011
ENA-CHECKLIST
Wallenberg Centre for Molecular Medicine, Linköping University, 58185 Linköping, Sweden Department of Biomedical and Clinical Sciences, Division of Molecular Medicine and Virology; Faculty of Medicine and Health Sciences, Linköping University, 58185 Linköping, Sweden
INSDC center name
public
INSDC status
ERS18601991
SRA accession
E-MTAB-13951:SMAD4_Both_1
Submitter Id
ArrayExpress
broker name
M170117
cell_line
melanocyte
cell_type
not collected
collection date
human
common name
Protocols: Cells were detached with 2 mM EDTA-PBS for 5 min and then centrifuged 250 g for 5 min. Cells were resuspended in 2 ml PBS and counted; 250,000 cells were taken per replicate (N = 2 per condition). M170117 cells were cultured in at 37°C, 5% CO2, and 95% humidity in RPMI medium (11875093, Thermo Scientific) supplemented with 10% Calf Bovine Serum (12133C, Sigma-Aldrich), 10 U/ml Penicillin-Streptomycin (15140122, ThermoFisher Scientific) and 4mM L-Glutamine (25030, ThermoFisher Scientific). Passages were performed with 2 mM EDTA-PBS (AM9260G, ThermoFisher Scientific).  Stimulations were performed with TGFβ1 (RnD Systems 7754-BH-005, Stock: 10 µg/mL in 4mM HCl + 0.1% BSA, 1:5000 dilution of stock), and/or MEKi (Trametinib, Stock: 50 µM Active Biochemicals - A-1258, 1:1000 dilution of stock), and/or DMSO 20 hours prior to harvest for CUT&RUN. CUT&RUN Low Volume Urea was performed as described in (36355069), using anti-SMAD4 (ABIN6972727, antibodies-online), anti-SMAD4 (D3R4N #46535, Cell Signaling Technology), and IgG negative control (ABIN101961, antibodies-online). Library preparation was performed as described in (36355069) using the KAPA Hyper Prep Kit (Roche) and KAPA Duel Index Adapters.
description
adult
developmental stage
melanoma
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
skin of body
organism part
Homo sapiens
scientific_name
SMAD4_Both_1
title