GSM4633675: BMDMs polarized with INFG (Polysome-associated mRNA) replicate 2; Mus musculus; OTHER
Polysome-profiling was performed  using polarized BMDMs. A fraction (~10%) of the obtained cell population was lysed in TRIzol reagent (ThermoFisher Scientific) and used for RNA sequencing with total RNA as input. Remaining cells were lysed in a hypotonic lysis buffer, as described and and lysates were  applied onto a 5%-50% (w/v) sucrose gradient, fractionated, and fractions corresponding to mRNA associated more than 3 ribosomes were collected in TRIzol reagent for subsequent RNA extraction. Such pooled samples were used for RNA sequencing of efficiently translated polysome-associated mRNA. SmartSeq2 sequencing libraries were prepared