Generation and Characterization of Cell Lines

GFP-, GFP-TIA1b-, GFP-TIARb-, and GFP-HuR-expressing FT293 cell lines were generated using the Flp-In T-Rex System (Invitrogen) [17 (link)]. Immunocytochemistry was carried out as described [16 (link), 17 (link)]. Murine embryonic fibroblasts (MEFs) knocked-out for TIA1 [26 (link)], TIAR [27 (link)], or EIF2AK2 [32 (link)] were maintained as described [33 (link)]. The lentiviral particles were generated from plasmids of second generation (pLKO series) that were transfected into HEK293T and the supernatants were collected. The shRNA against the human EIF2AK2 gene comes from SIGMA MISSION (# TRCN0000196400).

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Carrascoso I., Alcalde J., Tabas-Madrid D., Oliveros J.C, & Izquierdo J.M. (2018). Transcriptome-wide analysis links the short-term expression of the b isoforms of TIA proteins to protective proteostasis-mediated cell quiescence response. PLoS ONE, 13(12), e0208526.

Publication 2018

Flp-In T-Rex System

Cell lines Embryonic Fibroblasts Immunocytochemistry Murine Plasmids Shrna Tia1

Corresponding Organization : Consejo Superior de Investigaciones Científicas

Other organizations : Centro Nacional de Biotecnología

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Variable analysis

independent variables

Generation of GFP-, GFP-TIA1b-, GFP-TIARb-, and GFP-HuR-expressing FT293 cell lines using the Flp-In T-Rex System (Invitrogen)
Generation of murine embryonic fibroblasts (MEFs) knocked-out for TIA1, TIAR, or EIF2AK2
Transduction of cells with lentiviral particles containing shRNA against the human EIF2AK2 gene

dependent variables

Immunocytochemistry analysis of the generated cell lines

control variables

Maintenance of murine embryonic fibroblasts (MEFs) as described in [33 (link)]

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