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. 2021 Mar 2;53(3):369–383. doi: 10.1038/s12276-021-00570-6

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© The Author(s) 2021

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. 2 — a Western blotting analysis of the effect of KAT5 knockdown on MAP1LC3B lipidation after insulin withdrawal for 6 h. b Cell death rate following insulin withdrawal for 24 h in HCN cells transfected with control (siCon) or KAT5-targeting (siKAT5) siRNA (n = 3). c The interaction of endogenous ULK1 or ULK2 with GSK3B was analyzed by immunoprecipitation with anti-GSK3B antibody after insulin withdrawal for 6 h. d Schematic representation of the ULK1 domain structure and fragment constructs. e Analysis of the ULK1 regions responsible for GSK3B interaction. The indicated HA-ULK1 truncation mutants were expressed in HEK293 cells and precipitated with an anti-HA antibody, and coimmunoprecipitation with endogenous GSK3B was determined by western blotting. The blots shown are representative of at least three experiments with similar results. f HCN cells were fixed and analyzed by the proximity ligation assay with primary antibodies against GSK3B and the HA-tag. The cells were imaged by confocal microscopy. Scale bar, 10 μm. The graph represents the number of dots per cell following insulin withdrawal (n = 9 cells). ***P < 0.001. ns not significant.