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. 2021 Aug 7;40:248. doi: 10.1186/s13046-021-02049-8

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© The Author(s) 2021

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — HSPD1 knockdown blocks cell proliferation and clonogenic ability of NSCLC cells. A) Western blot analysis of HSPD1 protein levels in A549, H1299 and H460 cells upon infection with 3 independent shRNAs (#44, #45 and #48) targeting HSPD1 compared to scramble-infected cells (pLKO). β-Actin was used as loading control. B) Real-time proliferation curves of A549, H1299 and H460 infected with non-targeting pLKO or shHSPD1. The cells’ confluency plotted over time is shown. P-values are from two-way ANOVA. Points are averages of biological replicates ± SD. ** < 0.01, **** < 0.0001. Colony formation of A549 (C) and H1299 (D) cells with pLKO or shHSPD1, stained with crystal-violet and quantified in triplicates. Bars are average of biological replicates ± SD. P-values are from unpaired t-test. ** < 0.01, *** < 0.001. FACS plots of A549 (E) and H1299 (F) cells infected with pLKO or shHSPD1 and stained with PI for cell cycle analysis. Bar graph showing the % of cells in each cell cycle phase of A549 (G) and H1299 (H) cells upon infection with pLKO or shHSPD1. P-values are from two-way ANOVA. Bars are average of biological replicates ± SD. * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001