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. 2014 Dec 1;25(24):3954–3961. doi: 10.1091/mbc.E14-01-0004

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© 2014 Zeng et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — LanCL2 interacts with Akt. (A) FLAG-tagged LanCL2 was transfected in HEK293 cells, and FLAG IP was carried out 24 h after transfection. Endogenous Akt was detected by Western blotting. (B) FLAG-tagged LanCL2 and HA-tagged Akt were coexpressed in HEK293 cells, and HA IP was carried out after 24 h; this was followed by Western analysis. (C and E) His pull-down assay was performed with purified His-LanCL2 protein and HepG2 cell lysates (C) or purified GST-Akt protein (E) and analyzed by Western blotting. (D) HepG2 cells were cotransfected with FLAG-LanCL2 and HA-Akt; this was followed by IP with FLAG antibody. (F) FLAG-tagged LanCL2 was coexpressed with different HA-tagged Akt deletion mutants and FLAG IP was carried out 48 h after transfection.