Generic sample from Homo sapiens
Homo sapiens
The THP-1 cell line was sub-cloned and one clone (#5) was selected for ability to differentiate relatively homogeneously in response to PMA. THP-1.5 was used for all subsequent experiments. THP-1.5 cells were cultured in RPMI, 10% FBS, Penicillin/Streptomycin, 10mM HEPES, 1mM Sodium Pyruvate, 50uM 2-Mercaptoethanol, and treated with 30ng/ml PMA (Sigma) to differentiate them into macrophage-like cells. PMA-differentiated THP-1.5 were stimulated with 10ng/ml LPS (Salmonella Minnesota, chromatographically purified, Sigma) over a time-course of 24h, in presence of PMA. Total cell lysates were harvested in TRIzol reagent. Undifferentiated and unstimulated, PMA-differentiated cells were harvested in TRIzol reagent at the beginning of the LPS time-course. Total RNA was purified from TRIzol lysates according to manufacturer's instructions.
THP-1 control, fraction 3
tag_length:38-48bp,target_rna_length:22-32bp