在肿瘤相关巨噬细胞中使用CUT&RUN试验研究组蛋白乳酸化的基因组图谱。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-04-11 DOI:10.1016/j.xpro.2025.103766

Alessandra De Leo, Alessio Ugolini, Filippo Veglia

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引用次数: 0

摘要

赖氨酸乳酸化是一种独特的组蛋白修饰，在表观遗传调控和基因转录中起着至关重要的作用。在这里，我们提出了一种方案，用于研究肿瘤相关巨噬细胞中组蛋白乳酸化的基因组图谱。我们描述了制备活细胞的步骤，通过荧光激活细胞分选（FACS）分离葡萄糖转运蛋白1+ (GLUT1+)单核细胞源性巨噬细胞（MDMs）的门控策略，结合乳酸特异性一抗，并使用qPCR定量DNA。该方案适用于肿瘤源性和体外生成的骨髓源性巨噬细胞。有关本协议使用和执行的完整细节，请参阅De Leo等人1。

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Protocol to study the genomic profile of histone lactylation with CUT&RUN assay in tumor-associated macrophages.

Lysine lactylation is a distinctive histone modification that plays a crucial role in epigenetic regulation and gene transcription. Here, we present a protocol for studying the genomic profile of histone lactylation with the CUT&RUN assay in tumor-associated macrophages. We describe steps for preparing live cells, gating strategies for isolating glucose transporter type 1 + (GLUT1+) monocyte-derived macrophages (MDMs) by fluorescence-activated cell sorting (FACS), binding lactyl-specific primary antibodies, and quantifying DNA using qPCR. This protocol is applicable to both tumor-derived and in-vitro-generated bone-marrow-derived macrophages. For complete details on the use and execution of this protocol, please refer to De Leo et al.¹.

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