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. 2021 Jan 29;4:127. doi: 10.1038/s42003-021-01647-8

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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 Parameters for mitochondrial activity and cellular energy status analyzed after control or FANCD2 siRNA transfection. n = 3 independent experiments. **p = 0.0015 (CI–CIII activity); ***p = 0.0004 (oxygen consumption); **p = 0.0017 (ATP synthesis); **p = 0.008 (ATP/AMP ratio). b RT-qPCR-based analysis of CFS gene expression in cells after control or FANCD2 siRNA transfection and treated or not with 20 mM NaN3 for 10 h. n = 5 (FHIT), n = 4 (WWOX), n = 3 (IMMP2L), n = 4 (PARK2) independent experiments. **p = 0.0017 (FHIT_siFANCD2), **p = 0.0052 (FHIT_siLacZ_NaN3), *p = 0.0491 (FHIT_siFANCD2_NaN3 vs. FHIT_siFANCD2), *p = 0.0288 (WWOX_siFANCD2), **p = 0.006 (WWOX_siLacZ_NaN3), ***p = 0.0002 (WWOX_siFANCD2_NaN3), ***p = 0.0005 (WWOX_siFANCD2_NaN3 vs. siFANCD2), **p = 0.0091 (IMMP2L_siFANCD2), *p = 0.0464 (IMMP2L_siLacZ_NaN3), **p = 0.0047 (PARK2_siFANCD2), ***p = 0.0002 (PARK2_siLacZ_NaN3), *p = 0.0184 (PARK2_siFANCD2_NaN3 vs. PARK2_siFANCD2). c RT-qPCR analysis of CFS gene expression after control or FANCD2 siRNA transfection of cells maintained at 20% or 3% oxygen. n = 5 (FHIT), n = 5 (WWOX), n = 3 (IMMP2L), n = 5 (PARK2) independent experiments. ****p < 0.0001 (FHIT_siFANCD2_20%), **p = 0.0019 (FHIT_siFANCD2_3%), *p = 0.011 (FHIT_siFANCD2_3% vs. FHIT_siFANCD2_20%), **p = 0.0013 (WWOX_siFANCD2_20%), ****p < 0.0001 (WWOX_siLacZ_3%), ****p < 0.0001 (WWOX_siFANCD2_3%), ****p < 0.0001 (WWOX_siFANCD2_3% vs. WWOX_siFANCD2_20%), ****p < 0.0001 (IMMP2L_siFANCD2_20%), ***p = 0.0008 (IMMP2L_siFANCD2_3%), **p = 0.0014 (IMMP2L_siFANCD2_3% vs. IMMP2L_siFANCD2_20%), **p = 0.006 (PARK2_siFANCD2_20%), *p = 0.0118 (PARK2_siLacZ_3%), *p = 0.0107 (PARK2_siFANCD2_3%). d Chromosome fragility in cells transfected with control or FANCD2 siRNA, treated with 0.3 µM APH and maintained at 3% or 20% oxygen. Left, example of a DAPI-stained metaphase spread; the arrows indicate a break. Right, total breaks are scored as the mean number of breaks per metaphase. A total of 196 (siLacZ_20%), 221 (siFANCD2 20%), 185 (siLacZ 3%), and 186 (siFANCD2_3%) metaphases were analyzed from n = 4 independent experiments. *p = 0.0166 (siFANCD2_20%), **p = 0.0053 (siFANCD2_3% vs. siFANCD2_20%). e Frequency of FRA6E breaks in cells maintained at 3% oxygen, transfected with control or FANCD2 siRNA and treated with 0.3 µM APH. A total of 119 (siLacZ_3%), and 104 (siFANCD2_3%) metaphases were analyzed from n = 3 independent experiments. Error bars are standard error of the mean (SEM).