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Related Experiment Video

Updated: Jan 17, 2026

Quick Fluorescent In Situ Hybridization Protocol for Xist RNA Combined with Immunofluorescence of Histone Modification in X-chromosome Inactivation
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Quick Fluorescent In Situ Hybridization Protocol for Xist RNA Combined with Immunofluorescence of Histone Modification in X-chromosome Inactivation

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Tissue-Specific and NF-kappaB-Independent Xist RNA Localization Patterns in Female Intestinal, Blood, and Muscle

Claudia D Lovell, Isabel Sierra, Emma M Welter

    Biorxiv : the Preprint Server for Biology
    |September 15, 2025
    PubMed
    Summary
    This summary is machine-generated.

    Female cells use diverse epigenetic strategies for X-chromosome inactivation (XCI), varying by tissue. This ensures proper gene expression while maintaining overall silencing, even with age.

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    Combined DNA-RNA Fluorescent In situ Hybridization FISH to Study X Chromosome Inactivation in Differentiated Female Mouse Embryonic Stem Cells
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    Combined DNA-RNA Fluorescent In situ Hybridization FISH to Study X Chromosome Inactivation in Differentiated Female Mouse Embryonic Stem Cells

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    Area of Science:

    • Genetics
    • Epigenetics
    • Cell Biology

    Background:

    • X-chromosome inactivation (XCI) equalizes gene expression between sexes via Xist RNA and epigenetic silencing.
    • Quiescent female lymphocytes lack typical XCI epigenetic marks, which are restored by NF-κB signaling upon activation.

    Purpose of the Study:

    • To investigate if XCI epigenetic phenotypes correlate with quiescence or NF-κB activation in diverse female progenitor and stem cells.
    • To understand tissue-specific epigenetic mechanisms governing XCI in somatic cells.

    Main Methods:

    • Analysis of Xist RNA patterns and epigenetic marks in progenitor and stem cells from intestine, blood, and muscle.
    • Correlation of Xist RNA localization with NF-κB activation status in different cell types.
    • Examination of Xist RNA dynamics and gene expression in muscle satellite cells with aging.

    Main Results:

    • Intestinal progenitors show variable Xist RNA patterns despite NF-κB activation.
    • Blood progenitors and neutrophils exhibit a strong link between NF-κB activation and Xist RNA localization.
    • Muscle satellite cells and myoblasts accumulate Xist RNA at the Xi independently of NF-κB activation, maintaining silent yet allowing specific gene expression.

    Conclusions:

    • Female somatic cells utilize diverse, tissue-specific epigenetic mechanisms for XCI.
    • These mechanisms enable cell-type-specific gene expression from the inactive X chromosome while preserving broad silencing.
    • Aging impacts Xist RNA localization in muscle stem cells, but functional silencing is maintained.