SAMN39465889
External Id
Center for Theoretical Problems in Physico-chemical Pharmacology
INSDC center name
2024-01-17T00:00:00Z
INSDC first public
2024-01-17T07:01:03.877Z
INSDC last update
live
INSDC status
Human
NCBI submission model
Human.1.0
NCBI submission package
ERS17767929
SRA accession
62
age
HaCaT
cell_line
epidermal keratinocytes
cell_subtype
keratinocytes
cell_type
not collected
collection_date
To silence FOSL1 in HaCaT cells, we produced lentiviral particles in HEK293 cells using lentiviral vector p17019250 (Evrogen, Russia) and a commercial packaging kit. The vector encoded shRNA directed to FOSL1. In parallel, we also assembled lentiviral particles that contained vector p17019250c (Evrogen, Russia) instead of p17019250 using the same protocol. Unlike p17019250, vector p17019250c encoded scrambled shRNA. In 72h, we collected and filtered the cell culture media that contained virions to transduce HaCaT cells, according to the manufacturers protocol. After transduction, the infected cells were grown in DMEM + 20 percent FBS supplemented with puromycin (3 ug per ml ) to get rid of non-transduced cells. The selection was monitored in a fluorescent microscope due the ability of transduced cells to express GFP. Then, we compared FOSL1 expression levels in transduced cells by qPCR using FOSL1-specific primers. We also assessed the reduction of FOSL1 protein by Western blot with FOSL1 specific antibodies (Cat. number 252421, Abcam, USA).
description
adult
dev_stage
normal
disease
not applicable
disease_stage
German
ethnicity
Russia: Moscow
geo_loc_name
healthy
health_state
not applicable
isolate
84
karyotype
Homo sapiens
http://purl.obolibrary.org/obo/NCBITaxon_9606
organism
not applicable
phenotype
caucasian
race
cell line
sample_type
male
sex
epidermis
tissue
lentivirally transfected HaCaT cells to silence the transcription factor FOSL1
title