title: GSM8248701: 8wk, BCG, donor3; Homo sapiens; RNA-Seq
library_construction_protocol: hNOG mice were euthanized by CO2 inhalation and bone marrow (BM) was collected from both femurs by flushing 10mL of PBS through the bones using a 25G needle. The marrow collected was passed through a 70µm strainer and red blood cells were lysed with 1mL ACK lysis buffer (Gibco) for 5min. Cells were washed with 10mL RPMI containing 2% FBS and centrifuged at 300g for 5min. The resulting cell pellet was resuspended in 1mL PBS and cells were counted using the trypan blue exclusion method. Live human CD45+ cells devoid of mature B, T and NK cells (hCD45+hCD3-hCD19-hCD20-hCD56-) were sorted on the BD FACS Aria IV. Single cell suspensions of hCD45+hCD3-hCD19-hCD20-hCD56- cells sorted from the BM of hNOG mice before treatment (week 0) and at 4 and 8 weeks post-AS01/BCG treatment were processed on the 10x Genomics Chromium platform using Next GEM Single Cell 5' Kit (10x Genomics) v1 or v1.1, targeting droplet encapsulation of up to 6,000 cells