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. 2012 Oct 29;109(46):18767–18772. doi: 10.1073/pnas.1203853109

Fig. 4.

Fig. 4.

eIF4E requirement of selectively translated DDR and survival mRNAs with eIF4G1 overexpression. (A) Cells silenced with NS or eIF4G1 shRNAs subjected to 8 Gy IR and fractionated by sucrose density gradient sedimentation into unbound mRNA, light (1–3) and heavy (≥4) ribosomes. (B) Immunoblot of NS and eIF4G1-silenced MCF10A cells at 12 h following 8 Gy IR from equal protein amounts of total cell lysates. (C) Relative rate of protein synthesis assessed by immunoprecipitation (IP) of de novo proteins shown following 2 h metabolic labeling with [35S]methionine with actinomycin D (20 µM) and MG132 (10 µM) at 2 h after 8 Gy IR. (D) Immunoblot of NS, eIF4E silenced or eIF4G1-silenced MCF10A cells following 8 Gy IR at 24 h, from equal protein amounts of total cell lysates. (E) Quantification of immunoblot studies shown in B. EMCV results derived from Renilla luciferase reporter studies (n = 3).