(A) Tri-chamber neuron culture is schematically represented (S: soma, M: middle/methocel, N: neurite). (B) Steady state levels of phosphorylated eIF2-alpha after CHX treatment of N-chamber axons. At indicated time points after CHX addition, both S- and N-chambers were lysed in dish and 25 μl of each sample were run on a 12% SDS gel and the membranes were stained with phosphorylated eIF2-alpha and beta-actin antibodies after Western blotting. (C) PRV GS443 retrograde infection in tri-chambers. CHX was added to N-chambers 1 h before infection. Images of S-chambers were taken 20 hpi (4x magnification). (D) S-chamber virus yields were calculated as pfu, 20 hpi in the absence or presence of CHX either in the N-/or S-chambers. Data are the mean ± SEM with**P<0.01 using one sample t-test. ND: not detected, n≥3.