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. 2019 Jul 17;9:10329. doi: 10.1038/s41598-019-46949-4

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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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Fractionation reveals p97 accumulation in specific cellular compartments. (A) MCF-7 and (B) MCF-10A cells were each plated for 24 hours and then treated with increasing amounts of CB-5083 (p97i). 24 hours after treatment, cells were collected and fractionated into cytoplasmic, soluble nuclear, and chromatin-bound fractions. Proteins from each fraction were then visualized by Western blot with the indicated antibodies. Actin and Histone H3 serve as loading controls for the indicated fractions. Full Western blot images are available in Fig. S5. (C–E) Relative quantitation of p97 accumulation in (C) cytoplasmic, (D) soluble nuclear, and (E) chromatin-bound fractions from three biological replicates. Values are normalized to total p97 levels (Fig. S4) and error bars represent +/− one standard deviation.