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PDAP1 RIP in UV-crosslinked human T cells

ABSTRACT: Human CD4+ Total memory cells (CD4+CD25-CD45RA-CCR7+/-) were activated for 5d with anti-CD3/anti-CD28 antibodies. About 40*10^6 cells were then irradiated twice with 254nm UV light at 0.2J and then lysed in RIPA buffer. 1mg of cell extract was incubated on a rotating wheel ~16 h at 4°C with anti-PDAP1 antibody. PDAP1:mRNA complexes were immunoprecipitated with protein-G dynabeads 4hrs at 4°C and then treated with proteinase K. RNA was then purified using TRI-reagent and RNA zymospin columns. Three independent donors were used for this experiment. Inputs were also sequenced. Total RNA was sequenced in paired-end mode. This experiment is assessing direct targets bound by PDAP1 in human T cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE183685 | GEO | 2022/01/10

REPOSITORIES: GEO

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