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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. 4 — A Knockdown of Pirh2 leads to an increase of HeLa cell viability after heat-shock exposition. Colony-formation assay of Pirh2 knockdown (Pirh2 KD), HuR knockdown (HuR KD), and control (scrambled) HeLa cells in normal condition (ctrl) and after 45 °C incubation for 3 h (HS). The test was performed in triplicates. B Statistical analysis of colony-formation assay. Error bars indicate ±SEM. C Western blot analysis demonstrating protein levels of Pirh2 and HuR in Pirh2 knockdown (Pirh2 KD), HuR knockdown (HuR KD), and control (scrambled) HeLa cells. D Pirh2 destabilizes HuR under heat shock. Western blot analysis of HuR level in HeLa cells with Pirh2 knockdown (Pirh2 KD) and control HeLa cells (scrambled) after incubating at 43 °C for 90 and 120 min. 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 for the corresponding time points according to Student’s t-test. E Pirh2 stabilizes c-Myc protein in HeLa cells. Western blot analysis of Pirh2, HuR, and c-Myc levels in HeLa cells with Pirh2 knockdown (Pirh2 KD) and control HeLa cells (scrambled), both in normal condition and after incubating at 43 °C for 90 min. 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 for the corresponding time points according to Student’s t-test.