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. 2021 Jun 8;11:665045. doi: 10.3389/fonc.2021.665045

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Copyright © 2021 Wang, Xia, Chen, Chen, Meng, Lv and Yang

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Knockdown .of PHLPP decrease sensitive to EGFR-TKI in EGFR sensitive cell line. (A) Knockdown efficiency of PHLPP in HCC827 cell line as determine by both at mRNA level and protein level using quantitative RT-PCR and immunoblotting, respectively (B) IC50 values (μM) were based on data obtained from growth inhibition assay. (C) Longer-term colony formation assay using to determine downregulation PHLPP expression enhances cell survival (D) Detection of apoptosis with annexin V/flow cytometry. (E) HCC827-shControl and HCC827-shPHLPP cell lines were exposed to indicated EGFR-TKIs for 24 h then harvested for detection of apoptosis with annexin V/flow cytometry. The apoptosis of % are mean ± SE of triplicate assay. (F) HCC827-shControl and HCC827-shPHLPP cell lines were treated with increase concentration gefitinib for 24 h. The cells were lysed and indicated protein were detected by western blotting.