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ovcar8 (CLS Cell Lines Service GmbH)

CLS Cell Lines Service GmbH ovcar8
$a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and <t>mRFP-OVCAR8</t> nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.$
Ovcar8, supplied by CLS Cell Lines Service GmbH, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ovcar-8+cells/OVCAR-8+Cells/pmc12480658-251-3-5
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ovcar8 - by Bioz Stars, 2026-09

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ovcar 8 cells (Creative Biolabs)

Creative Biolabs ovcar 8 cells
$a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and <t>mRFP-OVCAR8</t> nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.$
Ovcar 8 Cells, supplied by Creative Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human ovarian carcinoma cell line ovcar-8 (Creative Biolabs)

Creative Biolabs human ovarian carcinoma cell line ovcar-8
$a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and <t>mRFP-OVCAR8</t> nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.$
Human Ovarian Carcinoma Cell Line Ovcar 8, supplied by Creative Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human ovarian carcinoma cell line ovcar-8 - by Bioz Stars, 2026-09

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cv ovcar 8 iroa is ovcar 4 cell ovcar 8 cell ovcar 4 cell (IROA Technologies LLC)

IROA Technologies LLC cv ovcar 8 iroa is ovcar 4 cell ovcar 8 cell ovcar 4 cell
$a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and <t>mRFP-OVCAR8</t> nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.$
Cv Ovcar 8 Iroa Is Ovcar 4 Cell Ovcar 8 Cell Ovcar 4 Cell, supplied by IROA Technologies LLC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cv ovcar 8 iroa is ovcar 4 cell ovcar 8 cell ovcar 4 cell - by Bioz Stars, 2026-09

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ovcar 8 cell (IROA Technologies LLC)

IROA Technologies LLC ovcar 8 cell
$a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and <t>mRFP-OVCAR8</t> nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.$
Ovcar 8 Cell, supplied by IROA Technologies LLC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ovcar-8+cells/IROA+Internal+Standard+Kit/pmc11794426-118-49-52
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ovcar 8 cell - by Bioz Stars, 2026-09

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non small cell lung cancer nci h226 6 05 breast cancer t 47d 6 03 ovarian cancer ovcar (ATCC)

ATCC non small cell lung cancer nci h226 6 05 breast cancer t 47d 6 03 ovarian cancer ovcar
$a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and <t>mRFP-OVCAR8</t> nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.$
Non Small Cell Lung Cancer Nci H226 6 05 Breast Cancer T 47d 6 03 Ovarian Cancer Ovcar, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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non small cell lung cancer nci h226 6 05 breast cancer t 47d 6 03 ovarian cancer ovcar - by Bioz Stars, 2026-09

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cell lines u2os female atcc rrid cvcl 0042 ovcar 8 (ATCC)

ATCC cell lines u2os female atcc rrid cvcl 0042 ovcar 8

Figure 2. PARG inhibition causes PARP1 aggregation in response to laser-induced DNA damage (A) Western blot showing the overexpression of C-terminal GFP-tagged PARP1 in a <t>U2OS</t> PARP1/ cell line. (B) Schematic overview of live cell imaging to monitor PARP1 recruitment and dissociation in the PARP1-eGFP cell line. (C) Cells were pre-treated with 10 mM PARGi for 1 h and subjected to laser-induced damage. Images were taken every 30 s for 30 frames and representative images are shown at pre-bleach, and 1 min and 10 min post-bleach. Scale bar: 10 mm. (D) Bright spot intensity was quantiﬁed and normalized to maximum PARP1 intensity across 8 different experiments and data are presented as mean ± SD. ***p < 0.001 (Welch’s two-sample t test). (E) PARP1 signal area at the bright spot was quantiﬁed over time in the PARGi-treated condition. (F) PARP1 aggregates were quantiﬁed over time in the PARGi-treated and untreated conditions. (G) PARP1 aggregate size (in microns2) was plotted on the left y axis and aggregate intensity (in arbitrary units a.u.) was plotted on the right y axis over time. Once PARP1 aggregates are formed and detected, their size and intensity remain stable. See also Figure S3; Video S1.

Cell Lines U2os Female Atcc Rrid Cvcl 0042 Ovcar 8, supplied by ATCC, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cell lines u2os female atcc rrid cvcl 0042 ovcar 8 - by Bioz Stars, 2026-09

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cells adherent ovarian cancer cell lines ovcar 8 (ATCC)

ATCC cells adherent ovarian cancer cell lines ovcar 8

Cells Adherent Ovarian Cancer Cell Lines Ovcar 8, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cells adherent ovarian cancer cell lines ovcar 8 - by Bioz Stars, 2026-09

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adherent ovarian cancer cell lines ovcar 8 (ATCC)

ATCC adherent ovarian cancer cell lines ovcar 8

Combinatorial treatment effect of olaparib and GW in a translational approach using primary <t>OvCa</t> spheroids. Primary cells from two BRCA 1 (UF-364; UF-510) and one BRCA 2 mutated patient (UF-357) were grown as spheroids in ULA plates and analyzed for their response to olaparib and additional treatment either GI or GW. The solvent DMSO has been used as a control. (A) Relative (rel.) caspase activity was increased following treatment with olaparib and GW compared to olaparib mono-treatment. Data is presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. (B) Representative Brightfield (left) and CellTox Green (right) images of UF-357; UF-364 and UF-510 following the indicated treatment conditions, which generated the strongest combinatorial effect are displayed (UF-357: 0/100 µM olaparib; UF-364: 0/10 µM olaparib; UF-510: 0/200 µM olaparib). Scalebar: 250 μm.

Adherent Ovarian Cancer Cell Lines Ovcar 8, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ovcar-8 cell line (CEM Corporation)

CEM Corporation ovcar-8 cell line

Ovcar 8 Cell Line, supplied by CEM Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results

$a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and mRFP-OVCAR8 nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.$

Journal: Nature Communications

Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

doi: 10.1038/s41467-025-62296-7

Figure Lengend Snippet: a Schematic representation of adipose tissue-mimicking collagen-based organo-hydrogels (OHGs). b BODIPY (green) staining of human peritoneal adipose tissue and OHG (representative images from n = 7 patients and n = 3 OHGs). c Quantification of human peritoneal adipocyte ( n = 100 adipocytes/tissue from 7 patients) and silicone oil microdroplet ( n = 100 microdroplets from 3 OHGs) diameter. d Quantification of volume fraction occupied by oil in peritoneal adipose tissues ( n = 7 patients) and OHGs ( n = 3 gels). e Storage moduli of hydrogels and human peritoneal tissues ( n = 5 gels; n = 9 adipose and n = 5 connective tissues). f Normalised stress-relaxation curves of hydrogels and human peritoneal tissues ( n = 5 gels or tissues). g Spheroid area after 7 d relative to day 0 of multiple ovarian cancer cell lines ( n = 3 spheroids). h Spheroid area comparison of ovarian cancer cell lines after 7 d in collagen or OHGs relative to the collagen average ( n = 3 spheroids). i Collagen-I (grey) and mRFP-OVCAR8 nuclei (red) staining of OVCAR8 cells seeded on top of collagen or OHG at 25 μm gel depth after 7 d in culture (representative images from n = 3 experiments). j Quantification of hydrogel organotypic invasion of OVCAR8 cells after 7 d ( n = 3 experiments). Rhombuses indicate the average. k BODIPY (green), mRFP-OVCAR8 (red), and collagen I (grey) staining of peritoneal tissue explants and mRFP-OVCAR8 cells after 7 d in culture (representative images from n = 3 experiments). Arrowheads indicate invading mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells in peritoneal tissue explants and OHGs (42 μm depth) after 7 d culture (representative images from n = 3 experiments). n Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues or OHGs after 7 d ( n = 3 tissues from distinct donors or gels). Rhombuses indicate the average. For the data in ( c , d , g , h , j , l and n ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( e ). Error bars in ( e ) represent the s.e.m. Scale bars, 50 μm ( b , i , m ), 100 μm ( k ). Components of ( a ) have been created in BioRender. Gautrot, J. (2025) https://BioRender.com/rk3jchz . Source data are provided as a Source Data file.

Article Snippet: OVCAR3 (HTB-161, ATCC), OVCAR8 (305383, Cytion), OVCAR4 (SCC258, Sigma), Kuramochi (JCRB0098, JCRB) and CAOV3 (HTB-75, ATCC) cells were grown in RPMI medium, while Tyk-nu (JCRB0234.0, JCRB) and Tyk-nu.CPR (JCRB0234.1, JCRB) were cultured in Minimum Essential Medium (MEM).

Techniques: Staining, Comparison

a pFAK, pMLC, YAP/TAZ (green; left to right) and nuclei (blue) staining of OVCAR8 cells in collagen or organo-hydrogel (OHG) after 48 h in culture (representative images from n = 3 gels). b Quantification of immunofluorescence signal intensity of cytoplasmic pFAK, pMLC, relative to the average in collagen and cytoplasmic:nuclear ratio of YAP/TAZ in OVCAR8 cells embedded in collagen or OHG for 48 h ( n = 24 cells). c F-actin (yellow), nucleus (blue) and BODIPY (grey) staining of an OVCAR8 cell spread at the ECM-microdroplet interface. d Shape descriptors of OVCAR8 nuclei after 24 h in collagen or OHG ( n = 53 nuclei). e Relative spheroid area of OVCAR8 after 7 d in OHG ( n = 10 spheroids). f BODIPY (green) and mRFP (red) staining of peritoneal adipose tissue explants and mRFP-OVCAR8 cells (49 μm into the tissue from the surface) after 7 d in culture (representative images from n = 3 experiments). g Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. h Nuclei (red) of OVCAR8 cells in OHGs after 7 d treatment (representative images from n = 14 spheroids). i Quantification of spheroid area relative to DMSO control of OVCAR8 cells in OHGs ( n = 14 spheroids). i F-actin (green) and nuclei staining of CAOV3 spheroids embedded in OHGs for 7 d (representative images from n = 6 spheroids). k Quantification of spheroid area in CAOV3 cells in OHGs after 7 d ( n = 6 spheroids). l F-actin (green) and nuclei (blue) staining of Kuramochi spheroids in collagen or OHG for 7 d with or without TGFβ (representative images from n = 3 spheroids). For the data in ( b , d , g , i and k ), a two-sided unpaired t test was performed. For data in ( e ), a one-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed. Scale bars, 25 μm ( a , c ), 100 μm ( f , I ), 200 μm ( h , j ). Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

doi: 10.1038/s41467-025-62296-7

Figure Lengend Snippet: a pFAK, pMLC, YAP/TAZ (green; left to right) and nuclei (blue) staining of OVCAR8 cells in collagen or organo-hydrogel (OHG) after 48 h in culture (representative images from n = 3 gels). b Quantification of immunofluorescence signal intensity of cytoplasmic pFAK, pMLC, relative to the average in collagen and cytoplasmic:nuclear ratio of YAP/TAZ in OVCAR8 cells embedded in collagen or OHG for 48 h ( n = 24 cells). c F-actin (yellow), nucleus (blue) and BODIPY (grey) staining of an OVCAR8 cell spread at the ECM-microdroplet interface. d Shape descriptors of OVCAR8 nuclei after 24 h in collagen or OHG ( n = 53 nuclei). e Relative spheroid area of OVCAR8 after 7 d in OHG ( n = 10 spheroids). f BODIPY (green) and mRFP (red) staining of peritoneal adipose tissue explants and mRFP-OVCAR8 cells (49 μm into the tissue from the surface) after 7 d in culture (representative images from n = 3 experiments). g Quantification of mRFP-OVCAR8 organotypic invasion into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. h Nuclei (red) of OVCAR8 cells in OHGs after 7 d treatment (representative images from n = 14 spheroids). i Quantification of spheroid area relative to DMSO control of OVCAR8 cells in OHGs ( n = 14 spheroids). i F-actin (green) and nuclei staining of CAOV3 spheroids embedded in OHGs for 7 d (representative images from n = 6 spheroids). k Quantification of spheroid area in CAOV3 cells in OHGs after 7 d ( n = 6 spheroids). l F-actin (green) and nuclei (blue) staining of Kuramochi spheroids in collagen or OHG for 7 d with or without TGFβ (representative images from n = 3 spheroids). For the data in ( b , d , g , i and k ), a two-sided unpaired t test was performed. For data in ( e ), a one-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed. Scale bars, 25 μm ( a , c ), 100 μm ( f , I ), 200 μm ( h , j ). Source data are provided as a Source Data file.

Techniques: Staining, Immunofluorescence, Control

a Schematic representation of norbornene-functionalised hyaluronic acid (NB-HA)-based organohydrogels (OHGs) presenting matrix metalloproteinase (MMP)-degradable crosslinking peptides and cell adhesion ligands. b Storage moduli of collagen- and NB-HA-based OHGs ( n = 5 collagen-based and n = 3 NB-HA-based OHGs; average ± s.e.m.). c Normalised stress-relaxation curves of collagen- and HA-NB OHG ( n = 5 collagen-based and n = 3 NB-HA-based OHGs). d F-actin (grey) and nuclei (red) in OVCAR8 cells embedded in collagen- or NB-HA-based OHGs for 7 d (representative images from n = 9 spheroids). e OVCAR8 spheroid area in collagen- or NB-HA-based OHGs relative to collagen-based OHG average ( n = 9 spheroids). f F-actin (grey) and nuclei (red) of OVCAR8 cells embedded in GFOGER- or RGD-presenting NB-HA OHG (representative images from n = 9 spheroids). g BODIPY (green), nuclei (red), and F-actin (grey) of OVCAR8 in GFOGER- or RGD-presenting NB-HA OHG (representative images from n = 9 spheroids). Arrowheads indicate cells in direct contact with oil microdroplets. h OVCAR8 spheroid area in GFOGER- or RGD-presenting NB-HA OHG after 7 d culture relative to GFOGER OHG average ( n = 9 spheroids). i F-actin (grey) and nuclei (red) in OVCAR8 cells embedded in collagen-based OHGs for 7 d (representative images from n = 8 spheroids). j OVCAR8 spheroid area in collagen-based OHGs after 7 d culture relative to DMSO control ( n = 8 spheroids). k BODIPY (green) and mRFP (red) staining of peritoneal adipose tissue explants and mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion depth into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m OVCAR8 spheroid area quantifications in MMP-cleavable or non-cleavable NB-HA OHG after 7 d culture relative to MMP-cleavable control ( n = 11 spheroids). For the data in ( b , e , h , j and l ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( m ). Error bars in ( b ) represent the s.e.m. Scale bars, 100 μm ( g , k ), 200 μm ( d , f , i ). Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

doi: 10.1038/s41467-025-62296-7

Figure Lengend Snippet: a Schematic representation of norbornene-functionalised hyaluronic acid (NB-HA)-based organohydrogels (OHGs) presenting matrix metalloproteinase (MMP)-degradable crosslinking peptides and cell adhesion ligands. b Storage moduli of collagen- and NB-HA-based OHGs ( n = 5 collagen-based and n = 3 NB-HA-based OHGs; average ± s.e.m.). c Normalised stress-relaxation curves of collagen- and HA-NB OHG ( n = 5 collagen-based and n = 3 NB-HA-based OHGs). d F-actin (grey) and nuclei (red) in OVCAR8 cells embedded in collagen- or NB-HA-based OHGs for 7 d (representative images from n = 9 spheroids). e OVCAR8 spheroid area in collagen- or NB-HA-based OHGs relative to collagen-based OHG average ( n = 9 spheroids). f F-actin (grey) and nuclei (red) of OVCAR8 cells embedded in GFOGER- or RGD-presenting NB-HA OHG (representative images from n = 9 spheroids). g BODIPY (green), nuclei (red), and F-actin (grey) of OVCAR8 in GFOGER- or RGD-presenting NB-HA OHG (representative images from n = 9 spheroids). Arrowheads indicate cells in direct contact with oil microdroplets. h OVCAR8 spheroid area in GFOGER- or RGD-presenting NB-HA OHG after 7 d culture relative to GFOGER OHG average ( n = 9 spheroids). i F-actin (grey) and nuclei (red) in OVCAR8 cells embedded in collagen-based OHGs for 7 d (representative images from n = 8 spheroids). j OVCAR8 spheroid area in collagen-based OHGs after 7 d culture relative to DMSO control ( n = 8 spheroids). k BODIPY (green) and mRFP (red) staining of peritoneal adipose tissue explants and mRFP-OVCAR8 cells. l Quantification of mRFP-OVCAR8 organotypic invasion depth into human peritoneal tissues after 7 d ( n = 3 tissues from distinct donors). Rhombuses indicate the average. m OVCAR8 spheroid area quantifications in MMP-cleavable or non-cleavable NB-HA OHG after 7 d culture relative to MMP-cleavable control ( n = 11 spheroids). For the data in ( b , e , h , j and l ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( m ). Error bars in ( b ) represent the s.e.m. Scale bars, 100 μm ( g , k ), 200 μm ( d , f , i ). Source data are provided as a Source Data file.

Techniques: Control, Staining

a F-actin (grey) and DAPI (blue) in OVCAR8 cells embedded in collagen or collagen-based organo-hydrogels (OHG) for 24 h (representative images from n = 3 gels). Microdroplet locations are indicated with an asterisk. b BODIPY (green), F-actin (grey) and nuclei (blue) staining of OVCAR8 cells embedded in collagen-based OHG (left; n = 3 gels) or BODIPY (green), mRFP (red) staining of mRFP-OVCAR8 cells invading into peritoneal adipose tissue (right; n = 3 tissues from 1 donor). Arrowheads indicate oil microdroplet (left) and adipocyte (right) deformations at the contact points with cells. c Quantification of OVCAR8 organotypic invasion depth into peritoneal adipose tissues after 7 d ( n = 3 tissues from 1 donor). Rhombuses indicate the average. d Schematic representation of the Sylgard 184 PDMS-based OGH. e F-actin (grey) and nuclei (red) staining of OVCAR8 spheroids embedded for 7 d in collagen-based OHGs prepared with PDMS microdroplets of varying stiffness (representative images from n = 10 spheroids). f OVCAR8 spheroid area after 7 d in collagen-based OHGs with PDMS microdroplets prepared with varying crosslinker percentage, relative to average area in 2 wt% microbead OHGs ( n = 10 spheroids). g Schematic representation of the norbornene-functionalised hyaluronic acid (NB-HA) OGH with norbornene-functionalised bovine serum albumin (NB-BSA) used for RGD presentation at the microdroplet surface. h F-actin (grey) and nuclei (red) of OVCAR8 spheroids after 7 d in NB-HA OHG with localised presentation of RGD (left). Relative OVCAR8 spheroid area in NB-HA OHG with localised RGD presentation after 7 d in culture ( n = 11 spheroids; top right). BODIPY (green), nuclei (red), and F-actin (grey) staining of OVCAR8 cells in NB-HA OHG presenting RGD on the microdroplet surface (bottom right). Arrowhead indicates a cell spreading at the microdroplet-HA interface. i Schematic representation of the system to modulate microdroplet interfacial mechanics. j F-actin (grey) and nuclei (red) of OVCAR8 spheroids after 7 d in NB-HA OHGs (microdroplet-restricted RGD presentation) with varying microdroplet interfacial modulus (representative images from n = 9 spheroids). k OVCAR8 spheroid area in NB-HA OHGs with microdroplet-presenting RGD and varying protein nanosheet interfacial mechanics after 7 d in culture, relative to BSA-only control ( n = 9 spheroids). For the data in ( c and h ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( f and k ). Scale bars, 25 μm ( a , b ), 50 μm ( h , bottom right), 200 μm ( e , h left, j ). Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

doi: 10.1038/s41467-025-62296-7

Figure Lengend Snippet: a F-actin (grey) and DAPI (blue) in OVCAR8 cells embedded in collagen or collagen-based organo-hydrogels (OHG) for 24 h (representative images from n = 3 gels). Microdroplet locations are indicated with an asterisk. b BODIPY (green), F-actin (grey) and nuclei (blue) staining of OVCAR8 cells embedded in collagen-based OHG (left; n = 3 gels) or BODIPY (green), mRFP (red) staining of mRFP-OVCAR8 cells invading into peritoneal adipose tissue (right; n = 3 tissues from 1 donor). Arrowheads indicate oil microdroplet (left) and adipocyte (right) deformations at the contact points with cells. c Quantification of OVCAR8 organotypic invasion depth into peritoneal adipose tissues after 7 d ( n = 3 tissues from 1 donor). Rhombuses indicate the average. d Schematic representation of the Sylgard 184 PDMS-based OGH. e F-actin (grey) and nuclei (red) staining of OVCAR8 spheroids embedded for 7 d in collagen-based OHGs prepared with PDMS microdroplets of varying stiffness (representative images from n = 10 spheroids). f OVCAR8 spheroid area after 7 d in collagen-based OHGs with PDMS microdroplets prepared with varying crosslinker percentage, relative to average area in 2 wt% microbead OHGs ( n = 10 spheroids). g Schematic representation of the norbornene-functionalised hyaluronic acid (NB-HA) OGH with norbornene-functionalised bovine serum albumin (NB-BSA) used for RGD presentation at the microdroplet surface. h F-actin (grey) and nuclei (red) of OVCAR8 spheroids after 7 d in NB-HA OHG with localised presentation of RGD (left). Relative OVCAR8 spheroid area in NB-HA OHG with localised RGD presentation after 7 d in culture ( n = 11 spheroids; top right). BODIPY (green), nuclei (red), and F-actin (grey) staining of OVCAR8 cells in NB-HA OHG presenting RGD on the microdroplet surface (bottom right). Arrowhead indicates a cell spreading at the microdroplet-HA interface. i Schematic representation of the system to modulate microdroplet interfacial mechanics. j F-actin (grey) and nuclei (red) of OVCAR8 spheroids after 7 d in NB-HA OHGs (microdroplet-restricted RGD presentation) with varying microdroplet interfacial modulus (representative images from n = 9 spheroids). k OVCAR8 spheroid area in NB-HA OHGs with microdroplet-presenting RGD and varying protein nanosheet interfacial mechanics after 7 d in culture, relative to BSA-only control ( n = 9 spheroids). For the data in ( c and h ), a two-sided unpaired t test was performed. One-way analysis of variance (ANOVA) with Tukey’s correction for multiple comparisons was performed for the data in ( f and k ). Scale bars, 25 μm ( a , b ), 50 μm ( h , bottom right), 200 μm ( e , h left, j ). Source data are provided as a Source Data file.

Techniques: Staining, Control

$a Schematic representation of the organo-hydrogels (OHGs) with varying microdroplet diameter and volume fraction. b Storage modulus of collagen-based OHG with distinct microdroplet size ( n = 4 gels; average ± s.e.m.). The dashed line shows the predicted trend. c Normalised stress-relaxation curves of collagen-based OHG with distinct microdroplet size ( n = 4 gels). d BODIPY (green) and F-actin (red) and nuclei (blue) staining of OVCAR8 cells after 7 d in collagen-based OHG of 70 or 25 μm diameter microdroplets (representative images from n = 13 spheroids). Arrowheads indicate cells at the invasive front. e Quantification of relative OVCAR8 spheroid area in collagen-based OHG of distinct emulsion percentage of volume fraction and microdroplet diameter after 7 d in culture ( n = 13 spheroids). f BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells after 7 d invasion into adipose peritoneal tissue explants (representative images from n = 3 explants per donor). g Correlation between patient average adipocyte diameter and invaded OVCAR8 cells after 7 d, at 42 μm tissue depth, relative to number of cells at the tissue surface ( n = 6 donors; average ± s.e.m.). The dashed line shows the predicted trend. h Quantification of interdroplet distance in OHG ( n = 100 microdroplets). i Quantification of the percentage of invasive front OVCAR8 cells in contact with microdroplets in OHG ( n = 121 cells in 3 gels). j Quantification of percentage of Ki67 + cells, immunofluorescence signal intensity of cytoplasmic pFAK and pMLC relative to collagen average, and nuclear:cytoplasmic ratio of YAP/TAZ in OVCAR8 cells embedded in OHG of 70 or 25 μm diameter microdroplets for 24 h ( n = 55 cells). k F-actin (green) and DAPI (blue) staining of CAOV3 spheroids embedded for 7 d in collagen-based 25 μm microdroplet OHG in the presence or absence of TGFβ. Arrowheads indicate invading cells (representative images from n = 6 spheroids). l Quantification of CAOV3 spheroid area relative to non-TGFβ-treated control after 7 d in culture ( n = 6 spheroids). m Schematic illustration of the cell force-dependent invasion of adipose tissue enabled by the anisotropic mechanics of OHGs and the generation of migration tracks at the ECM-adipocyte/microdroplet interface. Arrows indicate the direction of the force. For the data in ( h , i , j and l ), a two-sided unpaired t test was performed. Coefficient of determination and Pearson correlation (two-tailed test) were performed in ( g ) to determine the relationship between tissue adipocyte diameter and OVCAR8 invasion. Error bars in ( b ) represent the s.e.m. Scale bars, 50 μm (bottom panels in d , k ), 100 μm ( f ), 200 μm (top panels in d , k ). Source data are provided as a Source Data file.$

Journal: Nature Communications

Article Title: Biomimetic organo-hydrogels reveal the adipose tissue local mechanical anisotropy regulates ovarian cancer invasion

doi: 10.1038/s41467-025-62296-7

Figure Lengend Snippet: a Schematic representation of the organo-hydrogels (OHGs) with varying microdroplet diameter and volume fraction. b Storage modulus of collagen-based OHG with distinct microdroplet size ( n = 4 gels; average ± s.e.m.). The dashed line shows the predicted trend. c Normalised stress-relaxation curves of collagen-based OHG with distinct microdroplet size ( n = 4 gels). d BODIPY (green) and F-actin (red) and nuclei (blue) staining of OVCAR8 cells after 7 d in collagen-based OHG of 70 or 25 μm diameter microdroplets (representative images from n = 13 spheroids). Arrowheads indicate cells at the invasive front. e Quantification of relative OVCAR8 spheroid area in collagen-based OHG of distinct emulsion percentage of volume fraction and microdroplet diameter after 7 d in culture ( n = 13 spheroids). f BODIPY (green) and mRFP (red) staining of mRFP-OVCAR8 cells after 7 d invasion into adipose peritoneal tissue explants (representative images from n = 3 explants per donor). g Correlation between patient average adipocyte diameter and invaded OVCAR8 cells after 7 d, at 42 μm tissue depth, relative to number of cells at the tissue surface ( n = 6 donors; average ± s.e.m.). The dashed line shows the predicted trend. h Quantification of interdroplet distance in OHG ( n = 100 microdroplets). i Quantification of the percentage of invasive front OVCAR8 cells in contact with microdroplets in OHG ( n = 121 cells in 3 gels). j Quantification of percentage of Ki67 + cells, immunofluorescence signal intensity of cytoplasmic pFAK and pMLC relative to collagen average, and nuclear:cytoplasmic ratio of YAP/TAZ in OVCAR8 cells embedded in OHG of 70 or 25 μm diameter microdroplets for 24 h ( n = 55 cells). k F-actin (green) and DAPI (blue) staining of CAOV3 spheroids embedded for 7 d in collagen-based 25 μm microdroplet OHG in the presence or absence of TGFβ. Arrowheads indicate invading cells (representative images from n = 6 spheroids). l Quantification of CAOV3 spheroid area relative to non-TGFβ-treated control after 7 d in culture ( n = 6 spheroids). m Schematic illustration of the cell force-dependent invasion of adipose tissue enabled by the anisotropic mechanics of OHGs and the generation of migration tracks at the ECM-adipocyte/microdroplet interface. Arrows indicate the direction of the force. For the data in ( h , i , j and l ), a two-sided unpaired t test was performed. Coefficient of determination and Pearson correlation (two-tailed test) were performed in ( g ) to determine the relationship between tissue adipocyte diameter and OVCAR8 invasion. Error bars in ( b ) represent the s.e.m. Scale bars, 50 μm (bottom panels in d , k ), 100 μm ( f ), 200 μm (top panels in d , k ). Source data are provided as a Source Data file.

Techniques: Staining, Emulsion, Immunofluorescence, Control, Migration, Two Tailed Test

Journal: Structure (London, England : 1993)

Article Title: PARG inhibition induces nuclear aggregation of PARylated PARP1.

doi: 10.1016/j.str.2024.09.006

Figure Lengend Snippet: Figure 2. PARG inhibition causes PARP1 aggregation in response to laser-induced DNA damage (A) Western blot showing the overexpression of C-terminal GFP-tagged PARP1 in a U2OS PARP1/ cell line. (B) Schematic overview of live cell imaging to monitor PARP1 recruitment and dissociation in the PARP1-eGFP cell line. (C) Cells were pre-treated with 10 mM PARGi for 1 h and subjected to laser-induced damage. Images were taken every 30 s for 30 frames and representative images are shown at pre-bleach, and 1 min and 10 min post-bleach. Scale bar: 10 mm. (D) Bright spot intensity was quantiﬁed and normalized to maximum PARP1 intensity across 8 different experiments and data are presented as mean ± SD. ***p < 0.001 (Welch’s two-sample t test). (E) PARP1 signal area at the bright spot was quantiﬁed over time in the PARGi-treated condition. (F) PARP1 aggregates were quantiﬁed over time in the PARGi-treated and untreated conditions. (G) PARP1 aggregate size (in microns2) was plotted on the left y axis and aggregate intensity (in arbitrary units a.u.) was plotted on the right y axis over time. Once PARP1 aggregates are formed and detected, their size and intensity remain stable. See also Figure S3; Video S1.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-PARP1 Cell Signaling Technology Cat#9542; RRID: AB_2160739 Rabbit anti-PAR Millipore Cat#MABE1031; RRID: AB_2665467 Rabbit anti-SIRT6 Cell Signaling Technology Cat#12486; RRID: AB_2636969 Rabbit anti-Histone H3 GeneTex Cat#GTX122148; RRID: AB_10633308 Mouse anti-GAPDH Proteintech Cat#60004; RRID: AB_2737588 Mouse anti-PARG Millipore Cat#MABS61; RRID:AB_10806473 Mouse anti-PARP1 R&D Biosystems, discontinued Cat#4338-MC; RRID: AB_354246 Rabbit anti-XRCC1 Cell Signaling Technology Cat#2375; RRID: AB_2218471 Mouse anti-PAR Abcam Cat#ab14459; RRID: AB_301239 Rabbit anti-cleaved PARP (Asp214) Cell Signaling Technology Cat#5625; RRID: AB_10699459 Rabbit Anti-b-tubulin Cell Signaling Technology Cat#2146; RRID: AB_2210545 Anti-rabbit IgG HRP-linked Cell Signaling Technology Cat#7074; RRID: AB_2099233 Anti-mouse IgG HRP-linked Thermo Fisher Scientific Cat#31432; RRID: AB_228302 Mouse anti-phospho-Histone H2A.X (Ser139) Millipore Cat#05–636; RRID: AB_309864 Rabbit anti-53BP1 Novus Biologicals Cat#NB100-904; RRID: AB_10002714 Alexa Fluor 488 goat anti-mouse Thermo Fisher Scientific Cat#A-11001; RRID: AB_2534069 Alexa Fluor 555 goat anti-mouse Thermo Fisher Scientific Cat#A-21422; RRID: AB_2535844 Alexa Fluor 555 goat anti-rabbit Thermo Fisher Scientific Cat#A-21428; RRID: AB_2535849 Alexa Fluor 647 goat anti-rabbit Thermo Fisher Scientific Cat#A-21245; RRID: AB_2535813 Anti-phospho-Histone H2A.X (Ser139), Alexa Fluor 488 Conjugate Millipore Cat#05-636-AF488; RRID:N/A Bacterial and virus strains MAX Efficiency DH5a Competent Cells Invitrogen Cat#18258012 PARP1 exon 7-targeting lentivirus Dr. Gregory Breuer, Yale University N/A Chemicals, peptides, and recombinant proteins Dimethyl sulfoxide Sigma-Aldrich Cat#D4540 Methyl methanesulfonate Sigma-Aldrich Cat#129925 PDD 00017273 (PARGi) Tocris Cat#5952 Olaparib MedChem Express Cat#HY-10162 5-Bromo-20-Deoxyuridine Sigma-Aldrich Cat#B5002 Talazoparib Selleckchem Cat#S7048 Hydrogen peroxide Sigma-Aldrich Cat#H1009 Temozolomide Selleckchem Cat#S1237 Z-VAD-FMK Selleckchem Cat#S7023 Critical commercial assays Subcellular Fractionation Kit for Cultured Cells Thermo Fisher Scientific Cat#78840 Deposited data Original western blot images This study Mendeley Data https://doi.org/10.17632/ ktpzmk93yg.1 Experimental models: Cell lines U2OS (female) ATCC RRID:CVCL_0042 OVCAR-8 (female) DCTD Tumor Repository RRID:CVCL_1629 U2OS PARP1 / (female) This study N/A (Continued on next page) e1 Structure 32, 2083–2093.e1–e5, November 7, 2024

Techniques: Inhibition, Western Blot, Over Expression, Live Cell Imaging

Figure 4. PARP1 catalytic activity is required for the formation of PARP1 aggre- gates (A) Representative images of the validation of the U2OS PAR sensor cell line with 5 mM olaparib treatment. Upon olaparib treatment, no PAR sensor signal is detected 1 min after laser-induced damage. (B) Bright spot intensity was quantiﬁed and normalized to maximum PAR intensity across 4 different experiments, ***p < 0.001 (Welch’s two- sample t test). (C) Representative images of PARP1-eGFP, APLF- PBZ-mCherry, and the merged images taken pre- bleach, and 15 min and 60 min after laser-induced damage. Cells were pre-treated with 10 mM PARGi for 1 h. The co-localization of PARP1 with PAR and PAR with PARP1 aggregates were represented as percentages calculated using ImageJ with a co- localization tolerance of 1 pixel. Data are pre- sented as mean ± SD, n = 4 nuclei. Scale bar: 5 mm. (D) Schematic of full-length PARP1 protein showing the catalytic domain and the site of the point mu- tation. Western blot showing overexpression of PARP1 catalytic variants, E988D, producing me- dium PAR chains, and E988A, producing no PAR chains in the PARP1/ cell line. The PARP1 cat- alytic variants show reduced PAR chain expression compared to PARP1 wildtype (WT). (E) Representative images of PARP1-WT, -E988A, and -E988D-eGFP cells pre-bleach and 7.5 min post-bleach after pre-treatment with 10 mM PARGi for 1 h. (F) Quantiﬁcation of normalized PARP1 intensity at the site of damage with and without PARGi treat- ment after laser-induced damage over 15 min. n = 3 nuclei, n.s. = not signiﬁcant (Welch’s two-sample t test). (G) Quantiﬁcation of PARP1 aggregates in PARP1- WT, -E988A, and -E988D-eGFP cell lines with and without PARGi treatment at 7.5 min after laser- induced damage. PARP1 aggregates form only in the PARP1 catalytically proﬁcient cell line. Data are presented as mean ± SD, n = 3 nuclei, *p < 0.05 (two-tailed unpaired t test). Scale bar: 10 mm. See also Figure S5.

Journal: Structure (London, England : 1993)

Article Title: PARG inhibition induces nuclear aggregation of PARylated PARP1.

doi: 10.1016/j.str.2024.09.006

Figure Lengend Snippet: Figure 4. PARP1 catalytic activity is required for the formation of PARP1 aggre- gates (A) Representative images of the validation of the U2OS PAR sensor cell line with 5 mM olaparib treatment. Upon olaparib treatment, no PAR sensor signal is detected 1 min after laser-induced damage. (B) Bright spot intensity was quantiﬁed and normalized to maximum PAR intensity across 4 different experiments, ***p < 0.001 (Welch’s two- sample t test). (C) Representative images of PARP1-eGFP, APLF- PBZ-mCherry, and the merged images taken pre- bleach, and 15 min and 60 min after laser-induced damage. Cells were pre-treated with 10 mM PARGi for 1 h. The co-localization of PARP1 with PAR and PAR with PARP1 aggregates were represented as percentages calculated using ImageJ with a co- localization tolerance of 1 pixel. Data are pre- sented as mean ± SD, n = 4 nuclei. Scale bar: 5 mm. (D) Schematic of full-length PARP1 protein showing the catalytic domain and the site of the point mu- tation. Western blot showing overexpression of PARP1 catalytic variants, E988D, producing me- dium PAR chains, and E988A, producing no PAR chains in the PARP1/ cell line. The PARP1 cat- alytic variants show reduced PAR chain expression compared to PARP1 wildtype (WT). (E) Representative images of PARP1-WT, -E988A, and -E988D-eGFP cells pre-bleach and 7.5 min post-bleach after pre-treatment with 10 mM PARGi for 1 h. (F) Quantiﬁcation of normalized PARP1 intensity at the site of damage with and without PARGi treat- ment after laser-induced damage over 15 min. n = 3 nuclei, n.s. = not signiﬁcant (Welch’s two-sample t test). (G) Quantiﬁcation of PARP1 aggregates in PARP1- WT, -E988A, and -E988D-eGFP cell lines with and without PARGi treatment at 7.5 min after laser- induced damage. PARP1 aggregates form only in the PARP1 catalytically proﬁcient cell line. Data are presented as mean ± SD, n = 3 nuclei, *p < 0.05 (two-tailed unpaired t test). Scale bar: 10 mm. See also Figure S5.

Techniques: Activity Assay, Biomarker Discovery, Western Blot, Over Expression, Expressing, Two Tailed Test

$Figure 5. PARP1 aggregates are associated with cell death responses (A) Representative images of U2OS PARP1-eGFP cells in response to the indicated damaging treatments. Cells were treated with 0.00125% MMS and 10 mM PARGi for 24 h. Diffuse cytoplasmic PARP1 signal (indicated with an asterisk) is observed only in cells with nuclear PARP1 aggregates. (B) Percentage of cells containing cytoplasmic PARP1 was quantiﬁed across the treatment conditions for 3 independent experiments. Data are presented as mean ± SD, n R 40 nuclei per condition, **p < 0.01 (two-tailed unpaired t test). (C) U2OS PARP1-eGFP cells were treated as indicated and the cytoplasmic and nuclear-soluble fractions were isolated. SIRT6 and b-tubulin represent the nuclear and cytoplasmic loading controls respectively. Cleaved PARP1 signal was observed in both the nuclear and cytoplasmic fractions in the PARGi and MMS co-treatment condition. In our plasmid, full-length PARP1 tagged with eGFP corresponds to approximately 141 kDa. After cleavage at the Asp214 site of PARP1, the larger fragment would be 141-24 = 117 kDa, which aligns with our results.$

Journal: Structure (London, England : 1993)

Article Title: PARG inhibition induces nuclear aggregation of PARylated PARP1.

doi: 10.1016/j.str.2024.09.006

Figure Lengend Snippet: Figure 5. PARP1 aggregates are associated with cell death responses (A) Representative images of U2OS PARP1-eGFP cells in response to the indicated damaging treatments. Cells were treated with 0.00125% MMS and 10 mM PARGi for 24 h. Diffuse cytoplasmic PARP1 signal (indicated with an asterisk) is observed only in cells with nuclear PARP1 aggregates. (B) Percentage of cells containing cytoplasmic PARP1 was quantiﬁed across the treatment conditions for 3 independent experiments. Data are presented as mean ± SD, n R 40 nuclei per condition, **p < 0.01 (two-tailed unpaired t test). (C) U2OS PARP1-eGFP cells were treated as indicated and the cytoplasmic and nuclear-soluble fractions were isolated. SIRT6 and b-tubulin represent the nuclear and cytoplasmic loading controls respectively. Cleaved PARP1 signal was observed in both the nuclear and cytoplasmic fractions in the PARGi and MMS co-treatment condition. In our plasmid, full-length PARP1 tagged with eGFP corresponds to approximately 141 kDa. After cleavage at the Asp214 site of PARP1, the larger fragment would be 141-24 = 117 kDa, which aligns with our results.

Techniques: Two Tailed Test, Isolation, Plasmid Preparation

Combinatorial treatment effect of olaparib and GW in a translational approach using primary OvCa spheroids. Primary cells from two BRCA 1 (UF-364; UF-510) and one BRCA 2 mutated patient (UF-357) were grown as spheroids in ULA plates and analyzed for their response to olaparib and additional treatment either GI or GW. The solvent DMSO has been used as a control. (A) Relative (rel.) caspase activity was increased following treatment with olaparib and GW compared to olaparib mono-treatment. Data is presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. (B) Representative Brightfield (left) and CellTox Green (right) images of UF-357; UF-364 and UF-510 following the indicated treatment conditions, which generated the strongest combinatorial effect are displayed (UF-357: 0/100 µM olaparib; UF-364: 0/10 µM olaparib; UF-510: 0/200 µM olaparib). Scalebar: 250 μm.

Journal: Scientific Reports

Article Title: Inhibiting ADAM17 enhances the efficacy of olaparib in ovarian cancer spheroids

doi: 10.1038/s41598-024-78442-y

Figure Lengend Snippet: Combinatorial treatment effect of olaparib and GW in a translational approach using primary OvCa spheroids. Primary cells from two BRCA 1 (UF-364; UF-510) and one BRCA 2 mutated patient (UF-357) were grown as spheroids in ULA plates and analyzed for their response to olaparib and additional treatment either GI or GW. The solvent DMSO has been used as a control. (A) Relative (rel.) caspase activity was increased following treatment with olaparib and GW compared to olaparib mono-treatment. Data is presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. (B) Representative Brightfield (left) and CellTox Green (right) images of UF-357; UF-364 and UF-510 following the indicated treatment conditions, which generated the strongest combinatorial effect are displayed (UF-357: 0/100 µM olaparib; UF-364: 0/10 µM olaparib; UF-510: 0/200 µM olaparib). Scalebar: 250 μm.

Article Snippet: Adherent ovarian cancer cell lines OVCAR-8 (RRID: CVCL_1629 ) , IGROV-1 (RRID: CVCL_1304) and SKOV-3 (RRID: CVCL_0532), purchased from American Type Culture Collection (ATCC) were grown with RPMI-1640 medium containing 10% fetal bovine serum (Gibco Life Technologies), 1% L-Glutamine (Sigma-Aldrich, St. Louis, MO, USA) and penicillin-streptomycin (3000 U pen./30,000 μg str. per 500 mL RPMI-1640; Biochrom).

Techniques: Solvent, Control, Activity Assay, CellTox Assay, Generated

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