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. 2021 Jun 5;12(6):581. doi: 10.1038/s41419-021-03871-w

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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 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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Fig. 5 — A Pirh2 upregulates c-Myc mRNA level in H1299 cells. qRT-PCR results of c-Myc expression for H1299 cells with stable Pirh2 overexpression (Pirh2 OE) and control H1299 cells (ctrl), both in normal conditions and after 16 h treatment with 1 μM doxorubicin. *p ≤ 0.01 according to Student’s t-test. B Pirh2 knockdown leads to a decrease of c-Myc mRNA level. qRT-PCR results of c-Myc expression for H1299 cells with stable Pirh2 knockdown (Pirh2 KD) and control H1299 cells (scrambled), both in normal conditions and after 16 h treatment with 1 μM doxorubicin. *p ≤ 0.01 according to Student’s t-test. C Pirh2 upregulates c-Myc protein level in H1299 cells. Western blot analysis of c-Myc protein level for H1299 cells with stable Pirh2 overexpression (Pirh2 OE) and control H1299 cells (ctrl), both in normal conditions and after 16 h treatment with 1 μM doxorubicin. D Pirh2 knockdown leads to a decrease of c-Myc protein level. Western blot analysis of c-Myc level in H1299 cells with stable Pirh2 knockdown (Pirh2 KD) and control H1299 cells (scrambled), both in normal conditions and after 16 h treatment with 1 μM doxorubicin. The normalized intensity ratios were calculated as ratios between the signals of proteins analyzed and the corresponding actin bands on the basis of three measurements. Error bars indicate ±SD. *p ≤ 0.05 vs. scrambled (vs. untreated for γH2AX) according to Student’s t-test. E, F Analysis of cell cycle phases distribution for H1299 cell lines with stable Pirh2 overexpression (Pirh2 OE) and control H1299 (ctrl) (left panel), and H1299 cells with stable Pirh2 knockdown (Pirh2 KD) and control H1299 cells (scrambled) (right panel), both in normal conditions and after 24 h treatment with 100 μM etoposide. The test was performed in triplicates. Error bars indicate ±SD. *p-value ≤ 0.01 according to Student’s t-test.