GSM8041640: LPM_RFP_Esr1_Mut_1; Mus musculus; RNA-Seq
The cells were double-sorted directly into 5 μl TCL (Qiagen) buffer containing 1% 2-mercaptoenthonol (Sigma). NR mutant LPMs were sorted by their corresponding gRNA-conjugated fluorescence reporter, as DAPI-CD19-CD3-Ly6C-CD11bhiF4/80hi from BMC mice at 4 weeks after immune reconstitution.   Total RNA was purified using RNAClean XP beads (Beckman Coulter). Polyadenylated mRNA was selected using an anchored oligo(dT) primer (5′–AAGCAGTGGTATCAACGCAGAGTACT30VN-3) and then reverse transcribed to cDNA. First-strand cDNA was subjected to limited PCR amplification followed by Tn5 transposon-based fragmentation using the Nextera XT DNA Library Preparation Kit (Illumina). Libraries were constructed following the Smart-seq2 RNA-seq library preparation protocol.