Protocol for automated multivariate quantitative-image-based cytometry analysis by fluorescence microscopy of asynchronous adherent cells

Besse, Laetitia; Rumiac, Typhaine; Reynaud-Angelin, Anne; Messaoudi, Cédric; Soler, Marie‐Noëlle; Lambert, Sarah; Pennaneach, Vincent

doi:10.1016/j.xpro.2023.102446

Cited by 3 publications

(2 citation statements)

References 43 publications

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“…CD16 downregulation was also verified using quantitative image based cytometry (QIBC), which has been rigorously applied in this context elsewhere ( Supplementary Figure S7A–C ). 18 As expected, CD49a was also upregulated on NK cells after treatment with TGF-β in vitro ( Supplementary Figure S4B ). CD16 expression was dramatically reduced on CD49a + cells, compared to CD49a neg , confirming that TGF-β induced mechanisms downregulate CD16 ( Supplementary Figure S4C and D ).…”

Section: Resultssupporting

confidence: 76%

See 1 more Smart Citation

TGF-β signalling limits effector function capacity of NK cell anti-tumour immunity in human bladder cancer

Wong,

McCulloch,

Alim

et al. 2024

eBioMedicine

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Section: Resultssupporting

confidence: 76%

“…QIBC analysis was based on Besse et al., 2023 18 with the following alterations. Firstly background subtraction of all channels (rolling ball method, 30-pixel size) was performed in ImageJ (version 1.53t).…”

Section: Methodsmentioning

confidence: 99%

TGF-β signalling limits effector function capacity of NK cell anti-tumour immunity in human bladder cancer

Wong,

McCulloch,

Alim

et al. 2024

eBioMedicine

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Protocol for automated multivariate quantitative-image-based cytometry analysis by fluorescence microscopy of asynchronous adherent cells

et al. 2023

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p53-dependent crosstalk between DNA replication integrity and redox metabolism mediated through a NRF2-PARP1 axis

Elfar,

Aning,

Ngai

et al. 2024

Nucleic Acids Research

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Mechanisms underlying p53-mediated protection of the replicating genome remain elusive, despite the quintessential role of p53 in maintaining genomic stability. Here, we uncover an unexpected function of p53 in curbing replication stress by limiting PARP1 activity and preventing the unscheduled degradation of deprotected stalled forks. We searched for p53-dependent factors and elucidated RRM2B as a prime factor. Deficiency in p53/RRM2B results in the activation of an NRF2 antioxidant transcriptional program, with a concomitant elevation in basal PARylation in cells. Dissecting the consequences of p53/RRM2B loss revealed a crosstalk between redox metabolism and genome integrity that is negotiated through a hitherto undescribed NRF2-PARP1 axis, and pinpoint G6PD as a primary oxidative stress-induced NRF2 target and activator of basal PARylation. This study elucidates how loss of p53 could be destabilizing for the replicating genome and, importantly, describes an unanticipated crosstalk between redox metabolism, PARP1 and p53 tumor suppressor pathway that is broadly relevant in cancers and can be leveraged therapeutically.

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Protocol for automated multivariate quantitative-image-based cytometry analysis by fluorescence microscopy of asynchronous adherent cells

Cited by 3 publications

References 43 publications

TGF-β signalling limits effector function capacity of NK cell anti-tumour immunity in human bladder cancer

TGF-β signalling limits effector function capacity of NK cell anti-tumour immunity in human bladder cancer

Protocol for automated multivariate quantitative-image-based cytometry analysis by fluorescence microscopy of asynchronous adherent cells

p53-dependent crosstalk between DNA replication integrity and redox metabolism mediated through a NRF2-PARP1 axis

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