THP-1_PMA_LPS_timecourse
RIKEN THP-1 short RNA time course data (PMA and LPS stimulation)
THP-1.5 cells were cultured 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. Two sets of mixed short RNA libraries were generated from THP-1.5 cells over time-courses of PMA differentiation (0, 2, 4, 12, 24h and 24, 48, 72, 96h ) and from PMA-differentiated THP-1.5 cells over a time-course of LPS stimulation (0, 2, 4, 12, 24h). Five unmixed short RNA libraries, each containing a specific range of short RNA lengths, were constructed from undifferentiated THP-1.5 cells. Unmixed Short RNA libraries were constructed as for the mixed library protocol: fraction1, 60~70bp; fraciton2, 70~80bp; fraction3, 80~90bp; fraciton4, 90~100bp; fraction5, 100~130bp.  Total RNA was extracted by AGPC (Acid-Guanidinium-Phenol-Chloroform) standard methods. Five individual samples were combined to make the mixed short RNA library. Two mixed library were made from PMA timecourse samples, and one from LPS timecourse samples. The short RNAs derived from each sample were tagged with a 4nt Tissue ID tag during the adaptor ligation step in order to distinguish them. Concatamerised tags derived from short RNAs were sequenced using the 454 sequencer (Roche) and Short RNA libraries were sequenced using 454 technology.