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. 2017 Mar 27;7:45385. doi: 10.1038/srep45385

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Copyright © 2017, The Author(s)

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(A) Silver-stained SDS-PAGE revealed the Beclin1-interacting proteins were immuno-isolated from NIH3T3 cells that stably expressed Flag-tagged Beclin 1 or vector using anti-Flag beads. The proteins in the gel bands were extracted and identified using mass spectrometry, and DNA topoisomerase IIβ were detected. (B) Exogenous co-immunoprecipitation detecting DNA topoisomerase IIβ and Beclin 1 in nuclear lysates of NIH3T3 cells that stably expressed Flag-tagged Beclin1 or vector. The samples were subjected to immunoprecipitation analysis using anti-Flag antibodies. TBP was used as an internal control for nuclear samples. (C) Exogenous immunoprecipitation detecting DNA topoisomerase IIβ in 293T cells that overexpressed Flag-tagged Beclin 1 and treated with or without IR. (D) Exogenous immunoprecipitation detecting DNA topoisomerase IIβ in NIH3T3 cells that overexpressed Flag-tagged Beclin 1 treated with or without IR. (E) Endogenous co-immunoprecipitation and immunoblotting analysis detecting DNA topoisomerase IIβ and Beclin 1 in Hela cells treated with or without IR. All data are representative of the results of three independent experiments. (Full-length gels are included in the Supplementary Information file).