Nanostring pan-Cancer Analysis of Transcript Sub-Polysomal vs. Polysomal Distribution in OV3121 Mouse Granulosa Cell Line

We tested whether treatment with Tumor Necrosis Factor Alpha (Tnfα) corresponded to mRNA transit from sub-polysome to polysomal fractions in a mouse GC model, OV3121 cells (DOI 10.1111/j.1349-7006.1995.tb03063.x). <br>Cells were treated with VEH or 10 ng/ml Tnfα for 3 or 12 hours, and clarified lysates were loaded onto 10-60% sucrose gradients in SW41 tubes in lysis buffer lacking Triton X-100. Gradients were prepared using a BioComp system and chilled to 4oC before use. Samples were ultracentrifuged in a SW41 rotor at 36,000 rpm for 3h and 10 min, at 4oC, then samples were fractionated using the BioComp system, monitoring absorbance at 260 nm while collecting fractions of approximately 0.4 mL each. Fractions were then stored at -80oC until use.<br>Ribosomal fractions prepared using were pooled into “SUB”-polysomal (80S through the two ribosome peak) and POLYsomal samples. (see DOI 10.1016/j.cell.2016.01.003) n=2 unique replicates per treatment were performed and run on a nCounter mouse PanCancer Pathways IO360 cartridge (#LBL-10545-01) was run on a Nanostring Sprint Profiler (Nanostring Technologies Inc. Seattle, WA) at the VA Medical Center Research Core Facility (Aurora, CO). Normalized Nanostring data are provided here, along with a separate Attributes file indicating sample identities, each in .csv format.<br>