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. 2018 Jul 20;28(8):833–854. doi: 10.1038/s41422-018-0065-z

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — REEP1/2- and EI24-mediated ER Shaping are Required for DNA Damage-induced Apoptosis. a Western blot (WB) analysis of lysates from HEK293 cells transfected with the vector or 3× Flag-EI24 and treated with DMSO, cpt, doxo or eto for 12 h. GAPDH served as a loading control. b Western blot analysis of lysates from wild-type (WT) or REEP1/2 double knockout (DKO) U2OS cells transfected with the indicated plasmids and treated with 1 μM doxo for 24 h. c Western blot analysis of lysates from wild-type and EI24-knockout (KO) U2OS cells treated with DMSO, 50 μM eto, 1 μM cpt or 1 μM doxo for 24 h. d Western blot analysis of wild-type and EI24-knockout U2OS cells transfected with 3× Flag-EI24 or 3× Flag-EI24ΔH1 and treated with 1 μM cpt for 24 h. e Quantification of flow cytometry data for wild-type, REEP1/2 double knockout (DKO), or EI24 knockout cells positive for Annexin V. Cells were treated with 1 μM cpt for 14 h. f Western blot analysis of U2OS cells overexpressing the indicated proteins and treated with 1 μM doxo for 24 h. g Western blot analysis of lysates from REEP1/2 double knockout U2OS cells transfected with the vector, REEP1 or REEP1ΔH2 and treated with 1 μM doxo for 24 h. h Western blot analysis of wild-type and EI24-knockout U2OS cells transfected with REEP1/2 and treated with 1 μM cpt for 24 h. i Western blot analysis of wild-type and EI24-knockout U2OS cells transfected with control (shControl) or REEP1/2 shRNA (shREEP1/2) and treated with 1 μM cpt for 24 h. j Western blot analysis of U2OS cells transfected with the indicated shRNA alone or together with 3× Flag-EI24 and treated with 1 μM cpt for 24 h