GSM9335507: Macrophages from Piezo1^flox/flox C57BL/6J mice following lipopolysaccharide (LPS) treatment-2; Mus musculus; RNA-Seq
Total RNA was extracted using Trizol reagent (thermofisher, 15596018) following the manufacturer's procedure. The total RNA quantity and purity were analysis of Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA, 5067-1511) , high-quality RNA samples with RIN number > 7.0 were used to construct sequencing library. After total RNA   was extracted, mRNA was purified from total RNA (5ug) using Dynabeads Oligo (dT) (Thermo Fisher, CA, USA) with two   rounds of purification.    Then the   cleaved RNA fragments were reverse-transcribed to create the cDNA by SuperScript™ II Reverse Transcriptase (Invitrogen,   cat.    1896649, USA), which were next used to synthesise U-labeled second-stranded DNAs with E. coli DNA polymerase I  (NEB, cat.   m0209, USA), RNase H (NEB, cat.   m0297, USA) and dUTP Solution (Thermo Fisher, cat.   R0133, USA).   The average insert size for the final   cDNA librarys were 300±50 bp.    At last, we performed the 2×150bp paired-end sequencing (PE150) on an Illumina  Novaseq™ 6000 following the vendor's recommended protocol.