GSM5738269: 210509Bha_D21-4593; Mus musculus; RNA-Seq
Single cell lung suspensions from Eml4-Alk mice administered QZ1 were stained with the following antibodies (clone, fluorophore, dilution): CD44 (IM7, BV605, 1:200), CD105 (MJ7/18, BV786, 1:200), Ly6-A/E (D7, PE, 1:200), CD11b (M1/70, APC-Cy7, 1:200), CD45 (30-F11, AF488, 1:400), and EpCAM (G8.8, PE-Cy7, 1:200). Cells were stained for 20 minutes, and DAPI (1:10,000) was added immediately prior to sort. FACS sorting was performed on a FACSAria II (BD). Flow cytometry data was analyzed by the FlowJo software (Treestar). The sort strategy is shown in Fig. S15 of Soleimany et al., bioRxiv 2021. At least 100,000 cells from each of the QZ1+ and QZ1- compartments were collected into RPMI-1640 + 2% heat-inactivated FBS and pelleted via centrifugation at 1800 rpm for 5 minutes. Cell pellets were lysed in Trizol (ThermoFisher), and RNA was extracted using RNEasy Mini Kits (Qiagen). Libraries were prepared using the Clontech SMARTer Stranded Total RNAseq Kit (Clontech) and precleaned.