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GCN2 knockout does not interfere with transgene reactivation in HeLa cells.

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posted on 2018-07-18, 17:45 authored by Annarosaria De Vito, Massimo Lazzaro, Ilaria Palmisano, Davide Cittaro, Michela Riba, Dejan Lazarevic, Makoto Bannai, Davide Gabellini, Maria Vittoria Schiaffino

(A) Immunoblotting of protein extracts from the HeLa-OA1 parental cell line and GCN2-KO clones 183#11, 185#5 and 239#1, immunodecorated with anti-GCN2 antibody. Arrow, GCN2 specific band. Ponceau staining was used as loading control. (B, C) Relative transgene (OA1) mRNA abundance in HeLa-OA1 cells and GCN2-KO clones, cultured in Met/Cys (B) or Thr (C) deprived medium for 24 h or 48 h, respectively, compared to full medium. Mean ± SD of 3 technical replicates from 1 experiment. Data are expressed as fold change vs. control (full medium = 1). Since independent clones may display variable reactivation responses (e.g. due to different levels of transgene expression in basal conditions), the results are not shown as means of the three clones, but as separate replicates.

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