GSM5738370: Eml4-Alk Lung; Mus musculus; RNA-Seq
Eml4-Alk mice (10–12 weeks post tumor induction) and age-matched C57BL/6 healthy controls (Jackson Labs; 18–22 weeks) were euthanized by isoflurane overdose, and lungs were excised and separated into lobes. For tumor-bearing lungs, tumors were separated from healthy tissue using a dissecting microscope. Tissue was minced, treated with digestion buffer (Hank's Balanced Salt Solution + 2% heat-inactivated FBS, supplemented with DNase and collagenase), and incubated at 37C for 30 minutes with rotation. Samples were filtered using a 70 μm filter and diluted with RPMI-1640 + 2% heat-inactivated FBS. Cell suspension was centrifuged at 1800 rpm for 5 minutes and the pellet was resuspended in ACK lysis buffer for 2 minutes, followed by quenching with buffer (PBS + 2% heat-inactivated FBS). Cell suspension was centrifuged and supernatant was discarded. CD45 cell depletion and viability enrichment was performed according to manufacturer's instructions (StemCell Technologies). Single cells were processed using the 10X Genomics Single Cell 3' platform using the Chromium Single Cell 3' Library & Gel Bead Kit V2 kit (10X Genomics), per manufacturer's protocol. 10X Genomics Single Cell 3' platform; Chromium Single Cell 3' Library & Gel Bead Kit V2