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(A) Cell synchronization to metaphase followed by <t>UV-mediated</t> <t>protein-RNA</t> crosslinking. (B) Sucrose gradient fractionation and pooling of low-density (LD) and high-density (HD) RNPs. (C) Immunoprecipitation (IP) of LD- and HD- RNP fractions using anti-hnRNPC Ab, followed by partial RNase digestion (dashed line). (D) Completion of the fCLIP procedure, small RNA cDNA library preparation, and next-generation sequencing <t>(NGS).</t>
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(A) Cell synchronization to metaphase followed by UV-mediated protein-RNA crosslinking. (B) Sucrose gradient fractionation and pooling of low-density (LD) and high-density (HD) RNPs. (C) Immunoprecipitation (IP) of LD- and HD- RNP fractions using anti-hnRNPC Ab, followed by partial RNase digestion (dashed line). (D) Completion of the fCLIP procedure, small RNA cDNA library preparation, and next-generation sequencing (NGS).

Journal: bioRxiv

Article Title: Repurposed hnRNPC binds mature mRNAs and safeguards the mitotic transcriptome

doi: 10.1101/2025.09.11.675670

Figure Lengend Snippet: (A) Cell synchronization to metaphase followed by UV-mediated protein-RNA crosslinking. (B) Sucrose gradient fractionation and pooling of low-density (LD) and high-density (HD) RNPs. (C) Immunoprecipitation (IP) of LD- and HD- RNP fractions using anti-hnRNPC Ab, followed by partial RNase digestion (dashed line). (D) Completion of the fCLIP procedure, small RNA cDNA library preparation, and next-generation sequencing (NGS).

Article Snippet: RNA was depleted from rRNA (NEB, E7405) and used for NGS library preparation (NEB, E7760), followed by Illumina sequencing (NextSeq2000, 100 bp, single read).

Techniques: Fractionation, Immunoprecipitation, cDNA Library Assay, Next-Generation Sequencing