GSM8513355: prostate epithelial cells, C_rep2; Mus musculus; RNA-Seq
Briefly, single cell suspension of mouse prostate tissue of wild type, L-gp130peKI/KI, PtenpeΔ/Δ and PtenpeΔ/Δ;L-gp130peKI/KI mice was done and magnetic cell separation (MagniSort technology, Thermo Fisher) was performed for EpCAM positive fraction using anti-mouse CD326 (EpCAM) Biotin antibody (13-5791-82, eBioscience). For higher RNA output, three wild type and three L-gp130peKI/KI mouse prostates, respectively, were pooled to generate one sample. Per genotype n≥5 samples were analyzed by RNA-Seq. RNA isolation was performed using TriReagent (T9424-100ML, Sigma-Aldrich) and ReliaPrep RNA Tissue Miniprep System (Z6111, Promega) according to the manufacturer's instructions. Library preparation was performed using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (E7760, New England Biolabs) according to manufacturer's instructions in combination with a poly(A) mRNA magnetic isolation module (E7490) and multiplex oligos for Illumina (E7600). Libraries were amplified with 11 PCR cycles and the library size was analyzed by Agilent Tape Station (G2938-90014, Agilent Technologies).