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. 2011 Oct 17;108(44):18102–18107. doi: 10.1073/pnas.1112635108

Fig. 4.

Fig. 4.

Silencing of eIF4G1 reduces protein synthesis in neurons and exacerbates OGD-induced death. (A) Neurons were transfected with either shRNA lentivirus targeting rat eIF4G1 (sh4G) or a nontargeting shRNA virus (nt) for 72 and 96 h. Cell lysates were analyzed by Western blot and probed to determine eIF4G1 protein level. eIF4G1 silencing was effective, but other translation initiation factors were not altered. (B) After 1 h OGD and 24 h reperfusion, protein synthesis was reduced in control (nt) and more so in sh4G cultures. Cotransfection of neurons for 72 h with human HA-eIF4G1 and sh4G (4G+sh4G) significantly restored protein synthesis relative to sh4G under control conditions and after 1 h OGD and 24 h reperfusion. (C) Viability was greater in 4G+sh4G cotransfected neurons compared with control (nt) and sh4G transfected neurons following OGD; the data parallel the changes in protein synthesis shown in B under the same conditions. (D) Western blot showed eIF4G1 silencing in sh4G transfected neurons and expression at greater than control levels in 4G+sh4G transfected neurons under control conditions (nt) and after 1 h OGD and 24 h reperfusion with little change in other initiation factors. Bars represent mean ± SD of n = 3–4 independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001.