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. Author manuscript; available in PMC: 2017 Dec 1.
Published in final edited form as: Amino Acids. 2016 Sep 10;48(12):2831–2842. doi: 10.1007/s00726-016-2318-9

Fig. 2.

Fig. 2

Upregulation of 4EBP1 mRNA and protein levels in GCN2(+/+), GCN2(−/−), eIF2α(S/S) and eIF2α(A/A) MEFs in response to essential amino acid deficiency. Cells were grown in sufficient medium (Suff) or in medium deficient in histidine (His-) methionine (Met-), leucine (Leu-) or arginine (Arg-). a-d) ATF4 mRNA levels after 12 h of culture in deficient medium, expressed as fold the mean value for cells cultured in complete medium, are shown for experiments in a) GCN2(+/+), b) GCN2(−/−), c) eIF2α(S/S) and d) eIF2α(A/A) MEFs. e,f) 4EBP1 protein abundance in cells after 24 h of culture, shown as e) representative western blots and f) bar graph of the abundance of 4EBP1 protein, expressed as fold of the level in the same cell type cultured in sufficient medium after normalizing values to β-tubulin. All values for mRNA and protein are means ± SEM for 3 separate experiments, with each experiment having 3 replicates. Bars not denoted by the same letter are significantly different by Student’s t-test comparison at p≤ 0.05.