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Fig. 2. Co-immunoprecipitations of eif6 and gipc2 and of eif6 and gipc2mut. a. We utilized lysates of embryos injected with pCS2gipc2GFP. IP experiments were performed using anti-GFP antibodies for immunoprecipitation and anti-gipc2 and anti-eif6 antibodies for western blots. In western blots of the immunoprecipitations, the eif6 protein was found in the sample immunoprecipitated with anti-GFP (lane 1) and GFP was found in the sample immunoprecipitated with anti-eif6 (lane 3), indicating that eif6 and gipc2 co-immunoprecipitate. Lane 2=embryo lysate. In control IP using unrelated IgG the gipc2GFP and the eif6 bands are absent (lane 4). Lane 5=only an anti-GFP immunopositive band is present in the immunoprecipitate derived from lysates of pCS2GFP-injected embryos. b. We utilized lysates of embryos injected with pCS2gipc2mutHA (gipc2mut = gipc2 with PDZ domain deletion) to perform the IP experiments using anti-HA and anti-eif6 antibodies. The blots indicate that while in the lysates both eif6 and gipc2mutHA are present (lane 2), this does not occur in the reciprocal IPs, i.e., the gipc2mutHA band is absent in anti-eif6 IP (lane 1), and the eif6 band is absent in the anti-HA IP (lane 3). Therefore, gipc2mutHA does not interact with eif6. Lane 4=control IP using unrelated IgG: no bands are present. Image published in: De Marco N et al. (2017) Copyright © 2017. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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