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Fig. 4
DrPKZ-A and DrPKZ-B interact with eIF2α. (A) Co-immunoprecipitation (co-IP) of co-expressed V5-tagged protein (3.1a, PKZ-A-3.1a or PKZ-B-3.1a) and eIF2α-Myc (Danio rerio eIF2α with a C-terminal Myc tag) in HEK293T cells. WB1 and WB2 show that all recombinant vectors are expressed normally. An IP was performed using mouse anti-V5 antibody followed by immunoblotting with a rabbit anti-Myc antibody. Compared to the control sample, eIF2α coprecipitated with DrPKZ-A and DrPKZ-B, as shown in WB3. (B) Reciprocal co-IP experiments showed that DrPKZ-A and DrPKZ-B interact with eIF2α. IP was performed using a rabbit anti-Myc antibody, followed by immunoblotting with a mouse anti-V5 antibody. All the recombinant vectors express normally in HEK293T cells (WB1 and WB2). When the lysates from the control cells groups (lanes 1 and 3) were precipitated with anti-Myc antibody, the corresponding V5-tagged DrPKZ-A or DrPKZ-B was not detected by anti-V5 antibody. However, when the lysates from cells co-transfected with eIF2α-Myc and PKZ-A-3.1 (lane 2) or from cells co-transfected with eIF2α-Myc and PKZ-B-3.1 (lane 4) were precipitated with anti-Myc antibody, V5-tagged DrPKZ-A or DrPKZ-B was detected by anti-V5 antibody. The asterisk “*” marks nonspecific bands.
Reprinted from Gene, 527(1), Liu, Z.Y., Jia, K.T., Li, C., Weng, S.P., Guo, C.J., and He, J.G., A truncated Danio rerio PKZ isoform functionally interacts with eIF2alpha and inhibits protein synthesis, 292-300, Copyright (2013) with permission from Elsevier. Full text @ Gene