GSM2059932: IL15_Polysomal_A _1; Homo sapiens; RNA-Seq
Cytosolic and polysome-associated RNA was collected from 2-4 million NK cells per donor and treatment  (as described in Gandin et al.,  J. Vis. Exp. 2014). Briefly, total cytosolic mRNA was extracted (10% of the resulting input was kept as “cytosolic RNA”) and sedimented on a 5%-50% sucrose gradient to separate mRNA according to the number of associated ribosomes as described. Gradient fractions were collected and UV absorbance at 254 nm was measured. Fractions corresponding to >2 ribosomes (“polysome-associated RNA”) were collected. RNA was extracted using Tri reagent (Sigma, manufacturer’s protocol) and fractions were pooled followed by a column-based cleanup using the RNeasy MinElute Cleanup kit (Qiagen). smartSeq2 RNA-sequencing libraries were prepared as described in Picelli et al., 2013 Nat Methods (from step 5)
GEO Accession
GSM2059932