MM.1S cells, DMSO treatment, 24hrs
MM.1S cells were maintained in RPMI-1640 supplemented with 20% heat-inactive fetal bovine serum (hiFBS) and antibiotics in humidified atmosphere with 5% CO2 at 37C The MM.1S cells were treated with dIRF4 at 2.5 uM for 8 or 24 hrs with 0.1% DMSO. RNA was harvested using TRIzol Reagent (Invitrogene). 5 ug of total RNA was used for the construction of sequencing libraries. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.