***README file Miglietta et al. 2024***
Data Set Title: “Mitochondrial Chaperonin MCJ Promotes Vulnerability to Ferroptosis of Chemoresistant Ovarian Cancer Cells – Dataset”
Data Set Authors:
Stefano Miglietta (1), https://orcid.org/0000-0002-7755-2028; Manuela Sollazzo (1), https://orcid.org/0000-0001-5451-0063; Iacopo Gherardi (1), https://orcid.org/0000-0001-8100-924X; Sara Milioni (1,2), https://orcid.org/0000-0003-0357-8142; Beatrice Cavina (2), https://orcid.org/0000-0001-5485-5360; Lorena Marchio (2), https://orcid.org/0000-0002-7522-6325; Monica De Luise (2), https://orcid.org/0000-0002-3486-3985; Camelia Alexandra Coada (2), https://orcid.org/0000-0001-8362-6639; Marco Fiorillo (3), https://orcid.org/0000-0002-8055-3259; Anna Myriam Perrone (2,4), https://orcid.org/0000-0003-3140-4772; Ivana Kurelac (2), https://orcid.org/0000-0002-8364-9985; Giuseppe Gasparre (2), https://orcid.org/0000-0003-1229-9006; Luisa Iommarini (1), https://orcid.org/0000-0002-6804-7302; Anna Maria Ghelli (1,5), https://orcid.org/0000-0003-0830-0000; Anna Maria Porcelli (1,6), https://orcid.org/0000-0002-6209-0692.
Data Set Contact Persons: Luisa Iommarini (1), https://orcid.org/0000-0002-6804-7302; luisa.iommarini2@unibo.it 
Authors affiliations:
(1) Department of Pharmacy and Biotechnology (FABIT), University of Bologna, Bologna, Italy; (2) Department of Medical and Surgical Sciences (DIMEC), University of Bologna, Bologna, Italy; (3) Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Arcavacata di Rende, Italy; (4) Division of Gynecologic Oncology, IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy; (5) IRCCS Istituto delle Scienze Neurologiche di Bologna, Programma di Neurogenetica, Bologna, Italy; (6) Interdepartmental Centre for Industrial Research 'Scienze della Vita e Tecnologie per la Salute', University of Bologna, Bologna, Italy.
Data Set License: this data set is distributed under a Creative Commons Attribution 4.0 International (CC BY 4.0) license, https://creativecommons.org/licenses/by/4.0/  
Publication Year: 2024
Project Info: MOnARCHY (Dissecting the Metabolic signature of epithelial OvARian cancer: a challenge to overcome CHemotherapY resistance), funded by Associazione Italiana Ricerca sul Cancro. Grant number IG2020-ID24494.
Data set Contents
The data set consists of a compressed folder, named Dataset_Miglietta_et_al_2024.zip, containing 3 main folders: 
•	Folder_1 contains data showing that high MCJ levels sensitize OC cells to CDDP and decrease their proliferative capability
•	Folder_2 contains data showing that high levels of MCJ trigger ferroptosis in CDDP-resistant OC cells
•	Folder_3 contains data showing that MCJ-dependent lipid peroxidation triggers ferroptotic CDDP sensitivity
Each folder contains numerical data used for the generation of graphs in .csv format and images in .tif or .png formats.
Data set Documentation
Abstract
MCJ is a mitochondrial Tim23-related co-chaperonin known for its role in regulating oxidative phosphorylation efficiency, oxidative stress response and lipid metabolism. Recently, it has been proposed that the loss of MCJ correlates with cisplatin (CDDP)-resistance onset in ovarian cancer (OC), suggesting this protein as a potential prognostic factor during OC progression. However, the molecular mechanisms through which MCJ contributes to CDDP response remains poorly investigated. The present dataset collect data regarding the association between MCJ levels and accumulation of lipid droplets, tumorigenic features and sensitivity to CDDP in OC cells. Moreover, this dataset provides information on lipid peroxidation and ferroptosis induction in cells overexpressing MCJ, in presence or absence of Ferrostatin 1.
Content of the files
***Folder_1: High MCJ levels sensitize OC cells to CDDP and decrease their proliferative capability***
File 1A.csv contains values of cell viability expressed in percentage (%) obtained with cell viability assay and used to calculate IC50 after 72h of CDDP treatment in CDDP-sensitive ovarian cancer cell lines (SKOV3, OC314 and A2780) and CDDP-resistant counterparts (SKOV3cis, OC314cis and A2780cis).
File 1B.csv contains DNAJC15 mRNA expression levels as fold change on HPRT levels evaluated by qRT-PCR in CDDP-sensitive ovarian cancer cell lines (SKOV3, OC314 and A2780) and CDDP-resistant counterparts (SKOV3cis, OC314cis and A2780cis).
File 1C_A2780cis.csv contains values of cell viability to calculate IC50 after 72h of CDDP treatment in MCJ-OE cells and their respective controls (mock).
File 1C_OC314.csv contains values of cell viability to calculate IC50 after 72h of CDDP treatment in MCJ-KD cells and their respective controls (scramble).
File 1C_SKOV3cis.csv contains values of cell viability to calculate IC50 after 72h of CDDP treatment in MCJ-OE cells and their respective controls (mock).
File 1D.csv contains SRB absorbance data relative to colony formation assay of MCJ-OE and -KD OC cell lines compared to their respective controls.
Folder 1D contains representative pictures (.tif) of colony formation assay of MCJ-OE and -KD OC cell lines (relative to file 1D.csv).
File 1E_OC314.csv contains volume data (µm^3) of 3D cultures derived from OC314 (MCJ-KD and scramble) cell lines.
File 1E_SKOV3cis.csv contains volume data (µm^3) of 3D cultures derived from SKOV3cis (MCJ-OE and mock) cell lines.
Folder 1E contains representative pictures (.png) of 3D cultures derived from and OC314 (MCJ-KD and scramble) or SKOV3cis (MCJ-OE and mock) cell lines (related to files 1E_OC314.csv and 1E_SKOV3cis.csv)
File S1A.csv contains data regarding DNAJC15 mRNA expression levels as fold change to HPRT levels measured by qRT-PCR after overexpression in A2780cis and SKOV3cis cells.
Folder S1B contains representative images (.tif) relative to Western Blot analysis of DDK-myc tag (overexpressed MCJ) and VDAC levels performed on mitochondrial fractions of MCJ-OE and mock A2780cis and SKOV3cis cells.
File S1C.csv contains regarding DNAJC15 mRNA expression levels as fold change to HPRT levels measured by qRT-PCR after silencing with shRNA in OC314 cells.

***Folder_2: High levels of MCJ trigger ferroptosis in CDDP-resistant OC cells***
File 2A.csv contains the values relative to quantification of apoptotic cells after 72 hours of growth in absence of treatments, upon CDDP (+CDDP) and staurosporine (+STS) treatments
Folder 2A contains representative images (.tif) relative to quantification of apoptotic cells after 72 hours of growth in absence of treatments or upon CDDP and STS treatments (related to file 2A.csv).
File 2B.csv contains the values of total iron levels (ng/µL).
File 2C.csv contains the values of Fe2+ and Fe3+ fractions (%).
File 2D.csv contains the values of densitometric evaluation of GPX4 levels in total protein extracts expressed as fold change relative to calreticulin levels.
Folder 2D contains representative images (.tif or .png) relative to Western Blot analysis of GPX4 and calreticulin levels in total protein extract (related to file 2D.csv).
File 2E.csv contains data of GPX4 enzymatic activity.
File 2F.csv contains the values of densitometric evaluation of 4-HNE protein-adducts in total protein extracts expressed as fold change relative to HSP-70 levels.
Folder 2F contains representative images (.tif or .png) relative to Western Blot analysis of 4-HNE protein-adducts and HSP-70 levels in total protein extract (related to file 2F.csv).
File 2G.csv contains the quantification of lipid droplets in MCJ-OE and -KD cell lines compared with mock and scramble referred to the number of nuclei.
Folder 2G contains representative images (.png) relative to the quantification of lipid droplets and nuclei in MCJ-OE and -KD cell lines compared with mock and scramble (related to file 2G.csv).

***Folder_3: MCJ-dependent lipid peroxidation triggers ferroptotic CDDP sensitivity ***
File 3A.csv contains the values of densitometric evaluation of 4-HNE protein-adducts in total protein extracts expressed as fold change relative to HSP-70 levels with and without ferrostatin-1 (Fer-1) treatment.
Folder 3A contains representative images (.tif or .png) relative to Western Blot analysis of 4-HNE protein-adducts and HSP-70 levels in total protein extract with and without ferrostatin-1 (Fer-1) treatment (related to file 3A.csv).
File 3B.csv contains values of viable MCJ-OE and mock cells (expressed as percentage of treated vs untreated cells) upon treatments with CDDP treatment and/or ferrostatin-1.
File 3C.csv contains data regarding the correlation between ferrostatin-1 treatment effect and CDDP concentrations in MCJ-OE cells. 
File 3D.csv contains the quantification of lipid droplets in MCJ-OE cell lines compared with mock referred to the number of nuclei in presence or absence of ferrostatin 1.
Folder 3E contains representative images (.tif or .png) of Western Blot analysis of 4-HNE protein-adducts with and without DGAT1/2 inhibitors (related to file S3C.csv).
File 3F_A2780cis.csv contains the values of cell viability expressed in percentage (%) obtained with cell viability assay of mock and MCJ-OE A2780cis cells upon treatments with DGAT1/2i alone, CDDP alone or their combination.
File 3F_SKOV3cis.csv contains the values of cell viability expressed in percentage (%) obtained with cell viability assay of mock and MCJ-OE SKOV3cis cells upon treatments with DGAT1/2i alone, CDDP alone or their combination.
File S2A.csv contains the values of cell viability of MCJ-OE and mock cells with and without ferrostatin-1 treatment.
Folder S2B contains representative images (.png) of lipid droplets and nuclei in MCJ-OE cells with and without ferrostatin-1 treatment (related to file 3D.csv).
File S3A.csv contains data regarding lipid droplets in A2780cis MCJ-OE cells with and without DGAT1/2i treatment referred to the number of nuclei.
Folder S3A contains representative images (.png) of lipid droplets and nuclei in A2780cis MCJ-OE cells with and without DGAT1/2i treatment (related to file S3A.csv).
File S3B.csv contains data regarding lipid droplets in SKOV3cis MCJ-OE cells with and without DGAT1/2i treatment referred to the number of nuclei.
Folder S3B contains representative images (.png) of lipid droplets and nuclei in SKOV3cis MCJ-OE cells with and without DGAT1/2i treatment (related to file S3B.csv).
File S3C.csv contains the data regarding densitometric determination of Western Blot analysis of 4-HNE protein-adducts with and without DGAT1/2 inhibitors (DGAT1/2i) treatments (related to Folder 3E).
File specifics
To create the .csv files, GraphPad Prism v.10” was used with the following specifics:
•	UNICODE (UFT-8) 65001
•	Field delimiter « ; » (semicolon)
Notes
List of cell lines
•	SKOV3: ovarian cancer (OC) cell line
•	OC314: ovarian cancer (OC) cell line
•	A2780: ovarian cancer (OC) cell line
•	SKOV3cis: CDDP-resistant SKOV3 cell line
•	OC314cis: CDDP-resistant OC314 cell line
•	A2780cis: CDDP-resistant A2780 cell line
•	MCJ-OE: OC cells in which MCJ was overexpressed
•	MCJ-KD: OC cells in which MCJ was knocked down
•	mock: OC cells used as control for MCJ-OE cells
•	scramble: OC cells used as control for MCJ-KD cells