GSM5990228: Mo-AM WT-2; Mus musculus; RNA-Seq
Resting TR-AMs (CD45+CD11chiSiglec-Fhi) from naïve mice and TR-AMs (CD45+CD64+Siglec-FhiCD11blo) and Mo-AMs (CD45+CD64+Siglec-FloCD11bhi) from bleomycin-treated mice were sorted using the BD FACSAria™ II cell sorter. Total RNA was extracted from purified AM cells using the miRNeasy Mini Kit (QIAGEN). cDNA library was generated using SMART-Seq v4 Ultra Low Input RNA Kit (Clontech). cDNA was pre-amplified by a thermal cycler (Applied Biosystem) using 12-14 cycles and sheered into 150-400 bp fragments using Bioruptor Pico sonicator (Diagenode). The DNA ends were repaired using End-It™ DNA End-Repair Kit (Epicentre). The end repaired DNA fragments were ligated to adaptors by T4 ligase (NEB) after a single “A” was added at 3' by Klenow Fragment (3'→5' Exo-) (NEB). MinElute Reaction Cleanup Kit (Qiagen) was used for DNA cleanup after each step. Illumina P5 and P7 primers with individual adaptors were used for final library amplification. The 150-400 bp final product was purified on a 2% E-gel (Thermo Fisher Scientific).