RNA-Seq of Peritoneal macrophages treated with LBT
TotalRNAwasextractedusingTrizolreagent(Invitrogen,CA,USA)followingthemanufacturer'sprocedure.ThetotalRNAquantityandpuritywereanalysisofBioanalyzer2100andRNA6000NanoLabChipKit(Agilent,CA,USA)withRINnumber>7.0.Approximately10ugoftotalRNArepresentingaspecificadiposetypewassubjectedtoisolatePoly(A)mRNAwithpoly-Toligoattachedmagneticbeads(Invitrogen).Followingpurification,thepoly(A)-orpoly(A)+RNAfractionsisfragmentedintosmallpiecesusingdivalentcationsunderelevatedtemperature.ThenthecleavedRNAfragmentswerereverse-transcribedtocreatethefinalcDNAlibraryinaccordancewiththeprotocolforthemRNA-Seqsamplepreparationkit(Illumina,SanDiego,USA),theaverageinsertsizeforthepaired-endlibrarieswas300bp(50bp).Andthenweperformedthepaired-endsequencingonanIlluminaNovaseq6000atthe(lc-bio,China)followingthevendor'srecommendedprotocol.