Dataset DOI: 10.5061/dryad.8gtht771b

The following dataset contains numerical data and unprocessed images used during the processing of experiments described in an article entitled “Endogenous p21 levels protect genomic stability by suppressing both excess and restrained nascent DNA synthesis,” which is currently published in an open-access source. 

The numerical values correspond to quantification of the following assays: DNA fiber spreading, quantitative real-time PCR, micronucleus assay, anaphase aberration assay, immunostaining and fluorescence detection, and DNA polymerase foci detection. They are consolidated into a single Excel file with multiple sheets. Each sheet corresponds to the data used for a specific experiment as indicated by its name (inspired in the processed figure reported in the article). The section titled "Description of the data and file structure" provides a detailed explanation of the experimental context for each Excel sheet.

Grey cells: variables as described below.

Blue cells: data sets used to generate the analysis associated with the title of each sheet. This means blue cells indicate the samples used to generate the indicated figure.

Description of the data and file structure

The files and their respective descriptions are listed below.

Files and variables

File: Source_data_Excel_file.xlsx

Description: The Excel file called "Source data Excel file" is a consolidated file containing raw data corresponding to quantification of assays as described: DNA fiber spreading, quantitative real-time PCR, micronucleus assay, anaphase aberration assay, immunostaining and fluorescence detection, and DNA polymerase foci detection. (Source_data_Excel_file.xlsx).

To properly analyze the Excel file, it is essential to keep in mind that:

1)      The Excel cells highlighted in light blue in each sheet of the Excel file indicate which part of the data from these raw panels was used to generate figures for the above-mentioned manuscript.

2)      All data are organized in Excel sheets named according to the figure numbers they correspond to (e.g., Fig. 1A).

3)      Each spreadsheet contains the data used to generate the final analysis of the described experiment. For each variable, the corresponding experiment (1, 2, or 3) is specified where applicable.

4)      The Excel file with raw data does not contain merged cells, images, frozen panes, comments, hyperlinks, formulas, filters, or other elements that could affect the analysis.

The numerical values are associated with the following assays, as described: DNA fiber spreading: DNA track length (µm). Quantitative real-time PCR: CT values (qPCR cycles) used to calculate mRNA expression levels. Micronucleus assay: Number of micronuclei relative to the total number of nuclei counted. Anaphase aberration assay: Number of anaphase aberrations (bridge or lagging/acentric) relative to the total number of anaphases analyzed. Immunostaining and fluorescence detection: Signal intensity expressed in Arbitrary Units or as a percentage of total nuclei quantified. DNA polymerase foci detection: Number of nuclei containing polymerase foci (we quantified V5-PrimPol or GFP-Pol k foci) relative to the total number of nuclei analyzed.

The sheets corresponding to quantifications for the DNA fiber spreading assay have the following labels:

Fig. 1A, Fig. 1B, Fig. 1D, Fig. 2B, Fig. 3B, Fig. 5C, Fig. 5D, Fig. 5E, Fig. 5F, Fig. 6D, Fig. 7C, Fig. 7F, Fig. S1B, Fig. S3A, Fig. S3B, Fig. S4A, Fig. S4B, Fig. S4C, Fig. S4D, Fig. S5B, Fig. S5C, Fig. S5E, and Fig. S5G.

The sheets corresponding to quantitative real-time PCR data have the following labels:

Fig. 2A, Fig. 5B, Fig. 7B, Fig. S5A, Fig. S5D, Fig. S5F, Fig. S9B, and Fig. S11A.

The sheets corresponding to micronucleus assay data have the following labels:

Fig. 4A, Fig. 7D, Fig. 7E, Fig. 7G, Fig. 7H, Fig. S8A, Fig. S8D, and Fig. S10A.

The Sheet corresponding to the anaphase aberration assay has the following label:

Fig. 4B.

The sheets corresponding to immunostaining and fluorescence detection have the following labels, which include information on the target protein for each immunofluorescence assay in parentheses:

Fig. 2H (RPA-foci per nuclei), Fig. 3C (BrdU intensity), Fig. 3D (BrdU positive cells), Fig. 3E (gH2AX intensity), Fig. 4C (gH2AX intensity), Fig. 4D (ultra-fine bridges by PICH protein positive), Fig. 4E (53BP1 nuclear bodies by 53BP1 focis in nuclei), Fig. S8B (gH2AX intensity), Fig. S8C (53BP1 nuclear bodies by 53BP1 focis in nuclei), Fig. S8E (gH2AX intensity), Fig. S10B (gH2AX intensity), Fig. S10D (gH2AX intensity), and Fig. S10E (gH2AX intensity).

The sheets corresponding to DNA polymerase foci detection have the following labels and include the specific DNA polymerase for which the number of foci per nucleus is quantified, indicated in parentheses:

Fig. 2E (V5-PrimPol foci), Fig. 2F (V5-PrimPol foci), Fig. 2G (V5-PrimPol foci), Fig. 3A (V5-PrimPol foci), Fig. 6A (V5-PrimPol foci), Fig. 6C (V5-PrimPol foci), Fig. S6B (GFP-Pol k), Fig. S6C (GFP-Pol k), Fig. S6D (V5-PrimPol foci), Fig. S6E (GFP-Pol k), and Fig. S11B (V5-PrimPol foci).

Variables (highlighted in grey boxes)

The variables contained in the aforementioned Excel file are listed below.

• siLuc corresponds to the control sample with the transduction of siRNA (100nM) against luciferase.
• sip21 1 low corresponds to the sample with the transduction of siRNA (10nM) against p21. 
• sip21 1 high corresponds to the sample with the transduction of siRNA (100nM) against p21. 
• sip21 2 corresponds to the sample with the transduction of other siRNA (Ambion, s416) (100nM) against p21. 
• Olaparib corresponds to the treatment for 24 hs with the drug olaparib 10 uM.
• siPol eta corresponds to the sample with the transduction of siRNA (100nM) against pol eta. 
• sip53 corresponds to the sample with the transduction of siRNA (100nM) against to p53. 
• siIota corresponds to the sample with the transduction of siRNA (100nM) against pol Iota. 
• siPrimPol corresponds to the sample with the transduction of siRNA (100nM) against PrimPol. 
• siPol k corresponds to the sample with the transduction of siRNA (100nM) against Pol k. 
• EV corresponds to the control sample with the transduction of the empty vector plasmid (Plasmid #17482, Addgene)
• p21 corresponds to the sample with the transduction of the expression vector CS2-p21.
• p21PipMut corresponds to the sample with the transduction of expression vector CS2-p21 with the M147A, D149A and F150A mutations which were shown to disrupt the PCNA/p21 interaction have three mutations in its CDK binding domain (W49R, F51S and D52A) which disrupt the CDK2/p21 interaction.
• p21KO or p21 -/- corresponds to the sample of a cell with a p21 knock out.
• p53KO or p53 -/- corresponds to the sample of a cell with a p53 knock out.
• Pol I KO corresponds to the sample of a cell with an ADN Pol Iota knock out. 

File: Source_Western_blot_file_Calzetta_Mansilla_et_al.pdf

Description: The PDF file called "Source Western blot file Calzetta Mansilla et al" contains all raw data corresponding to Western Blot images (Source_Western_blot_file_Calzetta_Mansilla_et_al.pdf).

This file contains raw Western blot images, with the specific regions of interest indicated by red boxes. To the left of each blot, the protein detected in each experiment is specified. In some cases, different exposure times of the membrane are also indicated. 

File: Source_Representative_Images_file_Calzetta_Mansilla_et_al.pdf

Description: The PDF file called "Source Representative Images file Calzetta Mansilla et al" contains raw data corresponding to Representative Images related to the Excel file (Source_Representative_Images_file_Calzetta_Mansilla_et_al.pdf).

This file contains the raw images, with specific regions selected for presentation indicated by red boxes.

Code/software

N/A

Access information

Other publicly accessible locations of the data:

• N/A

Data was derived from the following sources:

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