Facs aria 2 cell sorted

We used an Hb9::GFP reporter stem cell line previously described infected with the Ngn2 lentiviral constructs as described above.^{29 (link)} Briefly, cells were differentiated in 24 well plates and subjected to different patterning molecules. At each time point, cells were dissociated with Accutase (Gibco) as previously described, each replicate was frozen in Cryostor CS10 (STEMCELL Technologies). After all samples were collected, cells were thawed in separated tubes are resuspended in sorting buffer as described by others,^{39 (link)} The BD FACS Aria II cell sorted was used to quantify the percentage of Hb9:GFP⁺ cells in each sample after using DAPI signal to determine cell viability.