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. 2021 Oct 4;11:19667. doi: 10.1038/s41598-021-99267-z

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© The Author(s) 2021

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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The effects of p53 on sensitivity to EGFR-TKIs in PC-9 cells. Endogenous p53 was silenced using a Crisper/Cas9 knockout system in PC-9 cells. (a) The indicated protein levels were analyzed by immunoblotting. (b,c) Two gefitinib-resistant cells (PC-9/GR and PC-9/p53^KO/GR) were established as described in the Materials and Methods section. Cells were treated with the indicated doses of gefitinib for 72 h, and cell viability was determined using MTT assays. (d) Lentiviral constructs containing the negative control (NT) and EGFR shRNAs were introduced into the indicated cells, and EGFR suppression was confirmed by immunoblotting. Cell viability was measured by cell counting. (e) Cells were treated with the indicated doses of gefitinib for 6 h, and EGFR-related signaling proteins were analyzed by immunoblotting. (f) Cells were treated with the indicated doses of osimertinib for 72 h, and cell viability was determined using MTT assays. (g) EGFR mutations were measured by the PNAClamp™. ***p < 0.0005 compared with negative control shRNAs.