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. 2021 Jul 5;12:4126. doi: 10.1038/s41467-021-24341-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 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 Fluorescent image (left panel) with quantification (right panel) showing similar levels of γH2AX foci in siControl (red) and siDHX9 (blue) cells treated with 1 μM camptothecin for 2 h. b Fluorescent image (left panel) with quantification (right panel) showing formation of RPA foci in siControl (red) and siDHX9 (blue) cells treated with 1 μM camptothecin for 2 h then recovered without the drug for the indicated times. DHX9 defective cells are impaired in formation of RPA foci. c RPA foci are dependent on the helicase activity of DHX9 and are diminished in cells overexpressing siRNA resistant helicase GFP-DHX9 dead mutant. Cells were treated with 1 μM camptothecin for 2 h. Quantification of n cells (as indicated) from three pooled biologically independent experiments were performed in (a), (b), and (c). Mean and error bars for one standard deviation are shown. Data sets were shown to be significantly different using a one-way ANOVA test with Tukey’s post hoc analysis as described in Methods. (ns not significant, *p < 0.1, **p < 0.01, ***p < 0.001, and ****p < 0.0001). d Western blot showing reduced RPA in the chromatin (P1) fraction of cells treated with 1 μM camptothecin (2 h) and knocked down for DHX9. RPA in the cytoplasmic (S1) and nucleoplasm (S2) fractions is not affected by the knockdown of DHX9. Lamin A/C is shown as a control for nuclear fractionation. Source data are provided as a Source Data file.