hsap_xg_001-tumor-lr-l-mef-03-x
Homo sapiens
Protocols: One million SCC25-GFP-Luc cells mixed with; i) one million mouse (MEFs), ii) one million naked mole-rat fibroblasts or, iii) no fibroblasts. 100 Î¼l of cell mixtures were injected subcutaneously to both flanks of 8-week old female immunosuppressed NSG mice. Tumors were collected 28 days after injection SCC25 tumors at day 28 were homogenized in lysis buffer from the Qiagen AllPrep mini kit using Tissue Lyzer II (Qiagen). Total RNA was then isolated using Qiagen AllPrep mini kit (Qiagen) and DNase digested (AM1907, Invitrogen) according to manufactures instructions. RNA concentration was measured using a NanoDrop spectrophotometer(Thermo Fisher): RNA integrity was assessed using RNA ScreenTapes(Agilent Technologies). The RNA integrity number (RIN) was above 8.6 forall samples. 100 ng DNase treated RNA was used to generate ribo-minustreated total RNA stranded libraries using Illumina Stranded Total RNAPrep Ligation with Ribo-Zero Plus kit (Illumina) according to manufacturesinstructions.