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. 2019 Nov 28;10:5428. doi: 10.1038/s41467-019-13315-x

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© The Author(s) 2019

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 — PIM1 inhibits apoptosis through phosphorylation of BAD. a Expression of PIM1 in SH-SY5Y or CHLA-20 cells transduced with non-targeting (NT) or PIM1-targeting gRNA molecules, by RT-qPCR and immunoblot. RT-qPCR data were normalized to cells treated with non-targeting gRNA and show means ± s.d. of triplicate experiments. b 72-h dose–response assays of brigatinib or ceritinib-exposed SH-SY5Y or CHLA-20 cells transduced with NT or PIM1-targeting gRNA. Data points represent means ± s.d. of triplicate experiments. ED₅₀ values were compared by an unpaired Student’s t-test. c Immunoblot of known PIM1 targets and apoptotic mediators in SH-SY5Y and CHLA-20 cells transduced with NT- or PIM1-targeting gRNA, the latter leading to PIM1 overexpression. Data are representative of three independent experiments. Numbers below the blot represent the densitometric measurement of pBAD levels normalized to BAD. d Percentage of apoptotic (annexin V + /PI-) PIM1-overexpressing SH-SY5Y cells after 48-h of treatment with 1 µM brigatinib or ceritinib. Data points in the bar chart show means ± s.d. of triplicate experiments. p-values were calculated using an unpaired Student’s t-test. Source data for this figure are provided as a Source Data file.