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. 2022 Jul 15;12:12078. doi: 10.1038/s41598-022-15244-0

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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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HA-CPNs Downregulate the Expression Levels and Signaling Activity of CD44 in GBM. (A) U-251 MG cells were treated with HA-CPNs and/or vehicle control (HA-CPN vehicle) and subjected to flowcytometry analysis of the indicated TIC populations; (B) Assessment of the protein expression levels of CD44 and CD44 intracytoplasmic domain (CD44-ICD) via Western blotting in FACS derived HA-CPN + and HA-CPN- cells (top). Protein levels quantified using densitometry and shown as ratio to Actin in Integrated Density Values (IDV) (bottom, graphs); (C) mRNA expression of signature markers of diverse endocytotic mechanisms in U-251 MG cells treated with 24 and/or 48 HA-CPNs/cell relative to HA-CPN vehicle control (dotted line); (D) CD44 and CD44-ICD protein expression levels in U-251 MG cells treated with HA-CPNs and/or HA-CPN vehicle control in the presence of Endosidin 9 or vehicle control (left). Protein levels quantified using densitometry and shown as ratio to Actin in Integrated Density Values (IDV) (right); (E) Flowcytometry analysis of CD44 expression levels in the indicated GBM patient samples treated with HA-CPNs and/or HA-CPN vehicle control. Data shown as mean ± s.d, n = 3, *p < 0.05, **p < 0.01, ***p < 0.001; Student’s t-test. Original blotted membranes of Western can be found in Fig. S7.