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. 2018 Sep 21;4:98. doi: 10.1038/s41420-018-0104-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. 2 — a RT-PCR for Bnip3-FL and Bnip3ΔExon3 in the heart of PND10 rat pups treated with misoprostol for 7 days. b RT-PCR for Bnip3 isoforms comparing expression patterns between human and mouse cell lines. c Bnip3 splicing diagram showing BNIP3ΔExon2 and Bnip3ΔExon3, indicating the differences in splicing between human and mouse. d Amino acid sequence alignment for human BNIP3ΔExon2 and mouse Bnip3ΔExon3. e Immunoblot showing that a commercially available antibody targeted to the amino-terminus (N-Terminus) of BNIP3 (CST #44060) can detect overexpressed HA-BNIP3-FL and HA-BNIP3ΔExon2. f Immunoblot demonstrating the specificity of our custom antibody targeted to the amino-terminus (N-Terminus) of BNIP3, where the antibody is able to detect overexpressed HA-BNIP3-FL (Human), Myc-Bnip3-FL (Mouse), and HA- Bnip3ΔExon3 (Mouse), but not HA-BNIP3ΔExon2 (Human). g Immunoblot demonstrating the specificity of si-BNIP3ΔExon3 (indicated as si-ΔEx3), shown using our custom antibody targeted to the amino-terminus (N-Terminus) of Bnip3. h H9c2 cells were exposed to 1% hypoxia for 24-h, with and without misoprostol treatment (10 μM). Extracts were western blotted as indicated. i Linear motif diagram for Bnip3-FL, human BNIP3ΔExon2, and mouse Bnip3ΔExon3. j Model of Mus Bnip3-FL indicating locations of the transmembrane (TM) domain, atypical BH3 domain (aBH3), and LC3-Interacting region (LIR) present on this isoform. k, l Comparison of Mus Bnip3ΔExon3 and human BNIP3ΔExon2, indicating the locations of the predicted ER domains, WH2 domains, nuclear localization sequence (NLS), and the Mus Bnip3ΔExon3 specific LC3-Interacting region (LIR)