GSM5352423: wt_EC4_S52450; Mus musculus; RNA-Seq
Colon lamina propria macrophages (Cd45+, Ly6G-, SiglecF-, Cd24-, Cd11c+/-, Cd103-, Cd11b+, Ly6C-, Cd64+, MHCII+) and Cd44+ epithelial cells (Cd45-, EpCAM+, Cd44+) from Tsc2fl/fl and Tsc2Lyz2-cre mice were isolated as per manufacturer's instructions (MACS Miltenyi Biotec, 130-097-410) and FACS sorted at the FACS Core Unit of the Children's Cancer Research Institute (CCRI) of Vienna. For the generation of full-length cDNA, the Smart-seq2 protocol (Picelli, S. et al, Nat Protoc. 2014 Jan;9(1):171-81. doi:10.1038/nprot.2014.006) was used. The subsequent library preparation from the amplified cDNA was performed using the Nextera XT DNA library prep kit (Illumina). For sequencing, libraries were pooled, diluted and sequenced on Illumina HiSeq 3000/4000 using 50 bp single-read chemistry.