GSM6042843: mNFIXko_Gene_Expression; Mus musculus; RNA-Seq
BM was extracted from femurs, pelvic bones, tibias and spines of TAM-treated Nfixflox/floxRosa26CreERT2+/+ and Nfixflox/floxRosa26CreERT2+/T mice via crushing c-Kit+ cells were isolated via magnetic enrichment using anti-CD117 microbeads (Miltenyi Biotec, Carlsbad, CA) and an autoMACs magnetic cell separator (Miltenyi Biotec, Carlsbad, CA). For c-Kit+ cells, HSPC (Lineage-Sca-1+cKit+) and LT-HSC (Lineage-Sca-1+c-Kit+CD48-CD150+) isolation, cells were stained with anti-Lineage BV605 cocktail, anti-Sca-1-PerCPCy5.5 (E13-161.7), anti-c-Kit-APCe780 (2B8), anti-CD48-Alexa Fluor 700 (HM48-1), anti-CD150-PE-Cy7 (TC15-12F12.2). For CMP (Lineage-Sca-1-c-Kit+CD34+CD16/32med) isolation, cells were stained with anti-Lineage BV605 cocktail, anti-Sca-1-PerCP-Cy5.5 (E13-161.7), anti-c-Kit-APCe780 (2B8), anti-CD34-FITC (RAM34) and anti-CD16/32-Alexa Fluor 700 (2.4G2). In addition to FACS antibodies, cells were labeled with CITE-seq antibodies (see Supplementary Table S1). 4′,6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich, St. Louis, MO) was used for dead cell exclusion. All cell isolations were performed using cell sorting on FACSAria III (BD Biosciences, San Jose, CA). Post cell sorting, 7000 Lineage-cKit+ BM, 1000 HSPC, 1000 CMP, and 1000 LT-HSC were pooled together for 10x genomics