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. 2022 Aug 19;12:14173. doi: 10.1038/s41598-022-18448-6

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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NEU and PPCA Protein Expression in Response to Multiple Differentiation Agents. HL60 cells were cultured for 7 days in the presence of DMF, DMSO, RA, or medium alone, lysed, and the lysates, at 50 µg total cellular protein/lane, were processed for NEU1 (A), PPCA (C), NEU2 (E), and NEU3 (G) immunoblotting. For the NEU4 immunoblots, cells were harvested on day 3 (I). (A, C, E, G, and I), to control for protein loading and transfer, blots were stripped and reprobed for β-actin. IB, immunoblot; IB*, immunoblot after stripping. MW in kDa is indicated on the left. (B, D, F, H, and J), densitometric analyses of the blots in (A, C, E, G, and I), respectively. n for each experimental group indicated in each panel (B and D) or under each vertical bar (F, H, and J). Vertical bars represent mean ± SE NEU1, PPCA, NEU2, NEU3, or NEU4 signal normalized to β-actin signal in the same lane on the same stripped and reprobed blot. *, increased normalized PPCA, NEU2, or NEU4 signal in dHL60 cells compared to HL60 cells at p < 0.05. **, decreased normalized NEU1 signal in dHL60 cells compared to HL60 cells at p < 0.05. Each blot is representative of 3–7 independent experiments. Cropped immunoblot images are shown.